QIAGEN QIAstat-Dx User Manual

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QIAGEN GmbH, QIAGEN Strasse 1, D

September 2020

QIAstat-Dx® Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook)

Version 1

For

in vitro

Rx Only

diagnostic use under Emergency Use Authorization Only

-40724 Hilden

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Contents

Intended Use .............................................................................................................. 4

Summary and Explanation ........................................................................................... 6

QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge Description ............................ 6

Pathogen Information .................................................................................................. 8

Principle of the Procedure .......................................................................................... 10

Description of the process ................................................................................ 10

Sample collection and cartridge loading ........................................................... 11

Sample preparation, nucleic acid amplification and detection ............................. 13

Materials Provided .................................................................................................... 14

Kit contents .................................................................................................... 14

Materials Required but Not Provided........................................................................... 15

Warnings and Precautions ......................................................................................... 16

Safety information .......................................................................................... 16

Reagent Storage and Handling .................................................................................. 19

Specimen Handling, Storage and Preparation .............................................................. 19

Procedure ................................................................................................................ 20

Internal Control .............................................................................................. 20

Protocol: Transport medium liquid samples ........................................................ 21

Viewing results ............................................................................................... 32

Interpretation of Results .............................................................................................. 34

Internal Control interpretation ........................................................................... 34

Pathogen Result interpretation .......................................................................... 34

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Quality Control ......................................................................................................... 43

Limitations ................................................................................................................ 44

Conditions of Authorization for the Laboratory ............................................................. 47

Performance Characteristics ....................................................................................... 49

SARS-CoV-2 Target ......................................................................................... 49

Clinical performance ...................................................................................... 49

Analytical performance ................................................................................... 50

Additional Targets included in the QIAstat-Dx SARS-CoV-2 Panel ......................... 53

Clinical performance ...................................................................................... 53

Expected values ............................................................................................. 73

Analytical performance ................................................................................... 80

Appendices............................................................................................................ 111

Appendix A: Installing the Assay Definition File ................................................ 111

Appendix B: Glossary ................................................................................... 113

Appendix C: Disclaimer of warranties ............................................................. 115

Symbols ................................................................................................................. 116

Ordering Information .............................................................................................. 117

Handbook Revision History ...................................................................................... 119

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Intended Use

The QIAstat-Dx Respiratory SARS-CoV-2 Panel is a multiplexed nucleic acid real-time PCR test intended for the qualitative detection and differentiation of nucleic acid from multiple respiratory viral and bacterial organisms, including the SARS-CoV-2 virus, in nasopharyngeal swabs (NPS) eluted in universal transport media collected from patients suspected of COVID-19 by their healthcare provider. Testing is limited to laboratories certified under the Clinical Laboratory Improvement Amendments of 1988 (CLIA), 42 U.S.C. §263a, to perform high complexity and moderate complexity tests.

The QIAstat-Dx Respiratory SARS-CoV-2 Panel is intended for the detection and differentiation of nucleic acid from SARS-CoV-2 and the following organism types and subtypes: Adenovirus, Coronavirus 229E, Coronavirus HKU1, Coronavirus NL63, Coronavirus OC43, SARS-CoV-2, Human Metapneumovirus A+B, Influenza A, Influenza A H1, Influenza A H3, Influenza A H1N1/pdm09, Influenza B, Parainfluenza virus 1, Parainfluenza virus 2, Parainfluenza virus 3, Parainfluenza virus 4, Rhinovirus/Enterovirus, Respiratory Syncytial Virus A+B,

pertussis

Chlamydophila pneumoniae

, and

Mycoplasma pneumoniae

Bordetella

SARS-CoV-2 RNA and nucleic acids from the other respiratory viral and bacterial organisms identified by this test are generally detectable in nasopharyngeal swabs (NPS) during the acute phase of infection. Positive results are indicative of the presence of the identified microorganism, but do not rule out co-infection with other pathogens not detected by the test, or lower respiratory tract infection that is not detected by a nasopharyngeal swab. The agent detected may not be the definite cause of disease.

For SARS-CoV-2 positive specimens; clinical correlation with patient history and other diagnostic information is necessary to determine patient infection status. Laboratories within the United States and its territories are required to report all SARS-CoV-2 positive results to the appropriate public health authorities.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Negative results do not preclude SARS-CoV-2 infection and should not be used as the sole basis for treatment or other patient management decisions. Negative SARS-CoV-2 results must be combined with clinical observations, patient history, and epidemiological information. Negative results for other organisms identified by the test may require additional laboratory testing (eg, bacterial and viral culture, immunofluorescence and radiography) when evaluating a patient with possible respiratory tract infection.

Testing with the QIAstat Dx Respiratory SARS-CoV-2 Panel is intended for use by qualified and trained operators who are proficient in performing the tests using the QIAstat Dx Analyzer System. The QIAstat Dx Respiratory SARS-CoV-2 Panel is only for use under the Food and Drug Administration’s Emergency Use Authorization.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Summary and Explanation

QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge Description

The QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge is a disposable plastic device that allows performance of fully automated molecular assays for the detection of respiratory pathogens. The main features of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge include compatibility with nasopharyngeal swab in transport medium (liquid samples), hermetical containment of the pre-loaded reagents necessary for testing, and true walk-away operation. All sample preparation and assay testing steps are performed within the cartridge.

All reagents required for the complete execution of a test run are pre-loaded and self-contained in the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge. The user does not need to come in contact with and/or manipulate any reagents. During the test, reagents are handled within the cartridge in the Analytical Module of the QIAstat-Dx Analyzer 1.0 by pneumatically-operated microfluidics and make no direct contact with the actuators. The QIAstat-Dx Analyzer 1.0 houses air filters for both incoming and outgoing air, further safeguarding the environment. After testing, the cartridge stays hermetically closed at all times, greatly enhancing its safe disposal.

Within the cartridge, multiple steps are automatically performed in sequence using pneumatic pressure to transfer samples and fluids via the transfer chamber to their intended destinations.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Swab port

Main port

Traceability bar code

Reaction chambers

After the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge containing the sample is introduced into the QIAstat-Dx Analyzer 1.0, the following assay steps occur automatically:

 Resuspension of Internal Control

 Cell lysis using mechanical and/or chemical means

 Membrane-based nucleic acid purification

 Mixing of the purified nucleic acid with lyophilized master mix reagents

 Transfer of defined aliquots of eluate/master mix to different reaction chambers

 Performance of multiplex real-time RT-PCR testing within each reaction chamber.

Note: An increase in fluorescence, indicating detection of the target analyte, is detected directly within each reaction chamber.

Figure 1. Layout of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge and its features.

Note: The swab port is not used for the QIAstat-Dx Respiratory SARS-CoV-2 Panel assay.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Pathogen Information

Acute respiratory infections can be caused by a variety of pathogens, including bacteria and viruses, and generally present with nearly indistinguishable clinical signs and symptoms. The rapid and accurate determination of the presence or absence of potential causative agent(s) helps make timely decisions regarding treatment, hospital admission, infection control, and return of the patient to work and family. It may also greatly support improved antimicrobial stewardship and other important public health initiatives.

The QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge is a single-use cartridge that includes all reagents needed for nucleic acid extraction, nucleic acid amplification and detection of 22 bacteria and viruses (or their subtypes), including SARS-CoV-2 that cause respiratory symptoms. Testing requires a small sample volume and minimal hands-on time, and the results are available in approximately one hour.

The SARS-CoV-2 target in the QIAstat-Dx Respiratory SARS-CoV-2 Panel has been designed upon alignment of more than 170 genomic sequences available in public databases from the SARS-CoV-2, which was identified as the causative agent of the viral pneumonia (COVID-19) outbreak originated in Wuhan, Hubei, China. The SARS-CoV-2 in this panel targets 2 genes of the virus genome (Orf1b poly gene (Rdrp gene) and E genes) detected with the same fluorescence channel. The two gene targets are not differentiated and amplification of either or both gene targets leads to a fluorescence signal.

Pathogens (and subtypes) that can be detected and identified with the QIAstat-Dx Respiratory SARS-CoV-2 Panel are listed in Table 1 (next page).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 1. Pathogens detected by the QIAstat-Dx Respiratory SARS-CoV-2 Panel

Pathogen Classification (genome type)

Influenza A Orthomyxovirus (RNA)

Influenza A, subtype H1N1/2009/pdm09 Orthomyxovirus (RNA)

Influenza A subtype H1 Orthomyxovirus (RNA)

Influenza A subtype H3 Orthomyxovirus (RNA)

Influenza B Orthomyxovirus (RNA)

Coronavirus 229E Coronavirus (RNA)

Coronavirus HKU1 Coronavirus (RNA)

Coronavirus NL63 Coronavirus (RNA)

Coronavirus OC43 Coronavirus (RNA)

SARS-CoV-2 Coronavirus (RNA)

Parainfluenza virus 1 Paramyxovirus (RNA)

Parainfluenza virus 2 Paramyxovirus (RNA)

Parainfluenza virus 3 Paramyxovirus (RNA)

Parainfluenza virus 4 Paramyxovirus (RNA)

Respiratory Syncytial Virus A/B Paramyxovirus (RNA)

Human Metapneumovirus A/B Paramyxovirus (RNA)

Adenovirus Adenovirus (DNA)

Rhinovirus/Enterovirus Picornavirus (RNA)

Mycoplasma pneumoniae

Chlamydophila pneumoniae

Bordetella pertussis

Bacterium (DNA)

Note: Enterovirus and Rhinovirus are both detected, but not differentiated, with the QIAstat-Dx Respiratory SARS-CoV-2 Panel.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Principle of the Procedure

Description of the process

Diagnostic tests with the QIAstat-Dx Respiratory SARS-CoV-2 Panel are performed on the QIAstat-Dx Analyzer 1.0. All of the sample preparation and analysis steps are performed automatically by the QIAstat-Dx Analyzer 1.0. Samples are collected and loaded manually into the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge:

A transfer pipette provided with the test kit is used for dispensing transport medium liquid sample into the main port (Figure 2).

Figure 2. Dispensing transport medium liquid sample into the main port.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Sample collection and cartridge loading

The collection of samples and their subsequent loading into the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge should be performed by personnel trained in safe handling of biological samples.

The following steps are involved and must be executed by the user:

1. A nasopharyngeal swab sample is collected.

2. The nasopharyngeal swab is placed into transport medium.

3. The sample information is manually written on or a sample label is affixed to the top of a QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge.

4. Transport medium liquid sample is loaded manually into the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge.

300 μl of sample is transferred into the main port of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge using one of the included transfer pipettes.

Note: When loading transport medium liquid sample, the user performs a visual check of the sample inspection window (see image below) to confirm that the liquid sample has been loaded (Figure 3, next page).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Figure 3. Sample inspection window (blue arrow).

5. The sample bar code and QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge QR code are scanned in the QIAstat-Dx Analyzer 1.0.

6. The QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge is introduced into the QIAstat-Dx Analyzer 1.0.

7. The test is started on the QIAstat-Dx Analyzer 1.0.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Sample preparation, nucleic acid amplification and detection

The extraction, amplification, and detection of nucleic acids in the sample are performed automatically by the QIAstat-Dx Analyzer 1.0.

1. The liquid sample is homogenized and cells are lysed in the lysis chamber of the QIAstatDx Respiratory SARS-CoV-2 Panel Cartridge, which includes a rotor that turns at high speed.

2. Nucleic acids are purified from the lysed sample via binding to a silica membrane in the purification chamber of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge in the presence of chaotropic salts and alcohol.

3. The purified nucleic acids are eluted from the membrane in the purification chamber and are mixed with the lyophilized PCR chemistry in the dried-chemistry chamber of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge.

4. The mixture of sample and PCR reagents is dispensed into the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge PCR chambers, which contain lyophilized, assay-specific primers and probes.

5. The QIAstat-Dx Analyzer 1.0 creates the optimal temperature profiles to carry out effective multiplex real-time RT-PCR and performs real-time fluorescence measurements to generate amplification curves.

6. The QIAstat-Dx Analyzer 1.0 Software interprets the resulting data and process controls and delivers a test report.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Materials Provided

Kit contents

QIAstat-Dx Respiratory SARS-CoV-2 Panel Catalog no. Number of tests

QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge* 6

Transfer pipettes† 6

* 6 individually packaged cartridges containing all reagents needed for sample preparation and multiplex real-time RT-

PCR, plus Internal Control.

†

6 individually packaged transfer pipettes for dispensing liquid sample into the QIAstat-Dx Respiratory SARS-CoV-2

Panel Cartridge.

691223 6

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Materials Required but Not Provided

The QIAstat-Dx Respiratory SARS-CoV-2 Panel is designed for use with the QIAstat-Dx Analyzer

1.0. Before beginning a test, make sure the following are available:

 QIAstat-Dx Analyzer 1.0 (at least one Operational Module and one Analytical Module)

with software version 1.2 or higher



QIAstat-Dx Analyzer 1.0 User Manual

 QIAstat-Dx latest Assay Definition File software for Respiratory SARS-CoV-2 Panel

installed on the Operational Module

(for use with software version 1.2 or higher)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Warnings and Precautions

For

in vitro

diagnostic use under Emergency Use Authorization only.

The QIAstat-Dx Respiratory SARS-CoV-2 Panel is to be used by laboratory professionals trained in the use of QIAstat-Dx Analyzer 1.0.

This device is restricted to sale by or on the order of a physician, or to a clinical laboratory; its use is restricted to, by, or on the order of a physician.

Pertussis is a nationally notifiable infectious disease in the U.S. If detected, notify state and/or local health departments.

Laboratories are required to report all positive SARS-CoV-2 results to the appropriate public health authorities.

Bordetella pertussis

Safety information

When working with chemicals, always wear a suitable lab coat, disposable gloves, and protective goggles. For more information, consult the appropriate safety data sheets (SDSs). These are available online in PDF format at www.qiagen.com/safety where you can find, view and print the SDS for each QIAGEN kit and kit component.

Always wear appropriate personal protective equipment, including but not limited to disposable powder-free gloves, a lab coat, and protective eyewear. Protect skin, eyes, and mucus membranes. Change gloves often when handling samples.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Handle all samples, used cartridges and transfer pipettes as if they are capable of transmitting infectious agents. Always observe safety precautions as outlined in relevant guidelines, such

(CLSI)

as the Clinical and Laboratory Standards Institute

from Occupationally Acquired Infections; Approved Guideline

Protection of Laboratory Workers

(M29), or other appropriate

documents provided by:

 OSHA

 ACGIH

: Occupational Safety and Health Administration (United States of America)

: American Conference of Government Industrial Hygienists (United States of

America)

 COSHH: Control of Substances Hazardous to Health (United Kingdom)

Follow your institution’s safety procedures for handling biological samples. Dispose of samples, QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridges, and transfer pipettes according to the appropriate regulations.

The QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge is a closed, single-use device that contains all reagents needed for sample preparation and multiplex real-time RT-PCR within the QIAstat-Dx Analyzer 1.0. Do not use a QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge that is past its expiration date, appears damaged, or leaks fluid. Dispose of used or damaged cartridges in accordance with all national, state, and local health and safety regulations and laws.

Observe standard laboratory procedures for keeping the working area clean and contamination-free. Guidelines are outlined in publications such as the

Microbiological and Biomedical Laboratories

from the Centers for Disease Control and

Biosafety in

Prevention and the National Institutes of Health (https://www.cdc.gov/labs/BMBL.html).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Highly flammable liquid and vapour. Harmful if swallowed or if

ory tract. Keep away from

dust/fume/gas/mist/vapours/spray. Wear protective gloves/protective clothing/eye protection/face protection. Wear

Continue rinsing. IF exposed or concerned: Immediately call a

and keep comfortable for breathing.

The following hazard and precautionary statements apply to components of the QIAstat-Dx Respiratory SARS-CoV-2 Panel.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge

Contains: ethanol; guanidine hydrochloride; guanidine thiocyanate; isopropanol; proteinase K; t-Octylphenoxypolyethoxyethanol. Danger!

inhaled. May be harmful in contact with skin. Causes severe skin burns and eye damage. May cause allergy or asthma symptoms or breathing difficulties if inhaled. May cause drowsiness or dizziness. Harmful to aquatic life with long lasting effects. Contact with acids liberates very toxic gas. Corrosive to the respirat heat/sparks/open flames/hot surfaces. No smoking. Avoid breathing

respiratory protection. IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do.

POISON CENTER or doctor/ physician. Remove person to fresh air

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Reagent Storage and Handling

Store the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridges in a dry, clean storage space at room temperature (15–25°C). Do not remove the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridges or the transfer pipettes from their individual packaging until actual use. Under these conditions, QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridges can be stored until the expiration date printed on the individual packaging. The expiration date is also included in the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge bar code and is read by the QIAstat-Dx Analyzer 1.0 when the cartridge is inserted into the instrument to run a test.

Specimen Handling, Storage and Preparation

Nasopharyngeal samples should be collected and handled according to the manufacturer’s recommended procedures.

Recommended storage conditions for NPS (nasopharyngeal swab) resuspended in UTM specimens are listed below:

 Room temperature up to 4 hours at 15–25°C

 Refrigerated up to 3 days at 2–8°C

 Frozen up to 30 days at –15 to –25°C

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Procedure

Internal Control

The QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge includes a full process Internal Control which is titered MS2 bacteriophage. The MS2 bacteriophage is a single-stranded RNA virus that is included in the cartridge in dried form and is rehydrated upon sample loading. This Internal Control material verifies all steps of the analysis process, including sample resuspension/homogenization, lysis, nucleic acid purification, reverse transcription and PCR.

A positive signal for the Internal Control indicates that all processing steps performed by the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge were successful.

A negative signal of the Internal Control does not negate any positive results for detected and identified targets, but it does invalidate all negative results in the analysis. Therefore, the test should be repeated if the Internal Control signal is negative.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Protocol: Transport medium liquid samples

Sample collection, transport and storage

Collect nasopharyngeal swab samples according to the swab manufacturer’s recommended procedures and place the swab into Universal Transport Medium.

Loading a sample into the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge

1. Open the package of a QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge using the tear notches on the sides of the packaging (Figure 4).

IMPORTANT: After the package is open, sample should be introduced inside the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge and loaded into the QIAstat-Dx Analyzer 1.0 within 120 minutes.

Figure 4. Opening the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge.

2. Remove the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge from the packaging and position it so that the QR code on the label faces you.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

3. Manually write the sample information or place a sample information label on the top of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge. Make sure that the label is properly positioned and does not block the lid opening (Figure 5).

Figure 5. Sample information placement on top of QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge.

4. Open the sample lid of the main port on the front of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge (Figure 6).

Figure 6. Opening the sample lid of main port.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

5. Open the tube with the sample to be tested. Use the supplied transfer pipette to draw up fluid to the third fill line on the pipette (i.e., 300 μl) (Figure 7).

IMPORTANT: Take care to avoid drawing air into the pipette. If Copan

UTM®, Universal Transport Medium is used as transport medium take care not to aspirate any of the beads present in the tube. If air or beads are drawn into the pipette, carefully expel the sample fluid in the pipette back into the sample tube and draw up fluid again.

Figure 7. Drawing up sample into the supplied transfer pipette.

6. Carefully transfer 300 μl of sample volume into the main port of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge using the supplied single-use transfer pipette (Figure 8, next page).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Figure 8. Transferring sample to main port of QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge.

7. Firmly close the sample lid of the main port until it clicks (Figure 9).

Figure 9. Closing the sample lid of the main port.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

8. Visually confirm that the sample has been loaded by checking the sample inspection window of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge (Figure 10).

IMPORTANT: After the sample is placed inside the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge, the cartridge must be loaded into the QIAstat-Dx Analyzer 1.0 within 90 minutes.

Figure 10. Sample inspection window (blue arrow).

Starting the QIAstat-Dx Analyzer 1.0

9. Power ON the QIAstat-Dx Analyzer 1.0 using the On/Off button on the front of the instrument.

Note: The power switch on the back of the Analytical Module must be set in the “I” position. The QIAstat-Dx Analyzer 1.0 status indicators will turn blue.

10. Wait until the Main screen appears and the QIAstat-Dx Analyzer 1.0 status indicators turn green and stop blinking.

11. Log in to the QIAstat-Dx Analyzer 1.0 by entering the user name and password.

Note: The Login screen will appear if User Access Control is activated. If the User Access Control is disabled, no user name/password will be required and the Main screen will appear.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

12. If the Assay Definition File software has not been installed on the QIAstat-Dx Analyzer 1.0, follow the installation instructions prior to running the test (see “Appendix A: Installing the Assay Definition File”, page 111, for additional information).

Running a test

13. Press the Run Test button in the top right corner of the touchscreen of the QIAstat-Dx Analyzer 1.0.

14. When prompted, scan the sample ID bar code on the UTM tube containing the sample, or scan the specimen information bar code located on the top of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge (see step 3) using the integrated front bar code reader of the QIAstat-Dx Analyzer 1.0 (Figure 11).

Note: It is also possible to enter the sample ID using the virtual keyboard of the touchscreen by selecting the Sample ID field.

Note: Depending on the chosen system configuration, entering the patient ID may also be required at this point.

Note: Instructions from the QIAstat-Dx Analyzer 1.0 appear in the Instructions Bar at the bottom of the touchscreen.

Figure 11. Scanning sample ID bar code.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

15. When prompted, scan the bar code of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge to be used (Figure 12). The QIAstat-Dx Analyzer 1.0 automatically recognizes the assay to be run based on the cartridge bar code.

Note: The QIAstat-Dx Analyzer 1.0 will not accept QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridges with lapsed expiration dates, previously used cartridges, or cartridges for assays that have not been installed on the unit. An error message will be shown in these cases and the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge will be rejected. Refer to the

QIAstat-Dx Analyzer 1.0 User Manual

for further details on how to install assays.

Figure 12. Scanning QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge bar code.

16. The Confirm screen will appear. Review the entered data and make any necessary changes by selecting the relevant fields on the touchscreen and editing the information.

17. Press Confirm when all the displayed data are correct. If needed, select the appropriate field to edit its content, or press Cancel to cancel the test (Figure 13, next page).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Figure 13. Confirming data entry.

18. Make sure that both sample lids of the swab port and main port of the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge are firmly closed. When the cartridge entrance port on the top of the QIAstat-Dx Analyzer 1.0 automatically opens, insert the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge with the bar code facing to the left and the reaction chambers facing down (Figure 14, next page).

Note: There is no need to push the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge into the QIAstat-Dx Analyzer 1.0. Position it correctly into the cartridge entrance port and the QIAstat-Dx Analyzer 1.0 will automatically move the cartridge into the Analytical Module.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Figure 14. Inserting QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge into QIAstat-Dx Analyzer 1.0.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

19. Upon detecting the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge, the QIAstat-Dx Analyzer 1.0 will automatically close the lid of the cartridge entrance port and start the test run. No further action from the operator is required to start the run.

Note: The QIAstat-Dx Analyzer 1.0 will not accept a QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge other than the one used and scanned during the test setup. If a cartridge other than the one scanned is inserted, an error will be generated and the cartridge will be automatically ejected.

Note: Up to this point, it is possible to cancel the test run by pressing the Cancel button in the bottom right corner of the touchscreen.

Note: Depending on the system configuration, the operator may be required to re-enter their user password to start the test run.

Note: The lid of the cartridge entrance port will close automatically after 30 seconds if a QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge is not positioned in the port. If this occurs, repeat the procedure starting with step 17.

20. While the test is running, the remaining run time is displayed on the touchscreen.

21. After the test run is completed, the Eject screen will appear (Figure 15, next page) and the Module status bar will display the test result as one of the following options:

 TEST COMPLETED: The test was completed successfully  TEST FAILED: An error occurred during the test  TEST CANCELED: The user canceled the test

IMPORTANT: If the test fails, refer to the “Troubleshooting” section in the

Analyzer 1.0 User Manual

for possible reasons and instructions on how to proceed.

QIAstat-Dx

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Figure 15. Eject screen display.

22. Press Eject on the touchscreen to remove the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge and dispose of it as biohazardous waste in accordance with all national, state, and local health and safety regulations and laws. The QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge should be removed when the cartridge entrance port opens and ejects the cartridge. If the cartridge is not removed after 30 seconds, it will automatically move back into the QIAstat-Dx Analyzer 1.0 and cartridge entrance port lid will close. If this occurs, press Eject to open the lid of the cartridge entrance port again and then remove the cartridge.

IMPORTANT: Used QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridges must be discarded. It is not possible to re-use cartridges for tests for which the execution was started but then subsequently cancelled by the operator, or for which an error was detected.

23. After the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge has been ejected, the results Summary screen will appear. Refer to “Interpretation of Results”, page, 34 for further details. To begin the process for running another test, press Run Test.

Note: For further information on the use of the QIAstat-Dx Analyzer 1.0, refer to the

QIAstat-Dx Analyzer 1.0 User Manual

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Viewing results

The QIAstat-Dx Analyzer 1.0 automatically interprets and saves test results. After ejecting the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge, the results Summary screen is automatically displayed (Figure 16).

Figure 16. Results Summary screen example showing Test Data on the left panel and Test Summary in the main panel.

The main part of the screen provides the following three lists and uses color-coding and symbols to indicate the results:

 The first list includes all pathogens detected and identified in the sample, preceded by a

sign and are colored red.

 The second list includes all equivocal pathogens, preceded by a yellow question

mark , in the event any of the subtypes H1, H3 and/or H1N1 pdm09 are detected and identified in the sample, but Influenza A is not detected.

 The third list includes all pathogens tested in the sample. Pathogens detected and

identified in the sample are preceded by a sign and are colored red. Pathogens that were tested but not detected are preceded by a sign and are colored green. Equivocal pathogens are preceded by a .

Note: Pathogens detected and identified in the sample are shown in all lists.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

If the test failed to complete successfully, a message will indicate “Failed”, followed by the specific Error Code.

The following Test Data is shown on the left side of the screen:

 Sample ID

 Assay Type

 Sample Type

Further data about the assay is available, depending on the operator’s access rights, through the tabs at the bottom of the screen (e.g., amplification plots and test details). For additional details, please see section below.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Interpretation of Results

Internal Control interpretation

Internal Control results are to be interpreted according to Table 2.

Table 2. Interpretation of Internal Control results

Control result Explanation Action

Passed The Internal Control amplified

successfully

Failed The Internal Control failed

Pathogen Result interpretation

The run was completed with success. All results are validated and can be reported. Detected pathogens are reported as “positive” and undetected pathogens are reported as “negative”.

Positively detected pathogen(s) are reported, but all negative results (tested but not detected pathogen[s]) are invalid.

Repeat the testing using a new QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge.

Result interpretation information for SARS-CoV-2:

The SARS-CoV-2 in this panel targets two genes of the virus genome (Orf1b poly gen (Rdrp gene) and E genes) detected with the same fluorescence channel. The two targets are not differentiated, and amplification of either or both regions leads to a fluorescence signal.

Result Interpretation information for Influenza A

A result for a respiratory organism is interpreted as “Positive” when the corresponding PCR assay is positive (see exceptions for Influenza A below). The Influenza A assay in the QIAstatDx Respiratory SARS-CoV-2 Panel is designed to detect Influenza A as well as Influenza A subtype H1N1/2009, Influenza A subtype H1, or Influenza A subtype H3. In particular, this means:

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

 If seasonal Influenza A H1 strain is detected by the QIAstat-Dx Respiratory SARS-CoV-2

Panel assay, two signals will be generated and displayed on the QIAstat-Dx Analyzer

1.0 screen: one for Influenza A and a second one for H1 strain.

Note: It is acceptable if only the H1 signal is obtained, which would be indicated as “equivocal”.

 If seasonal Influenza A H3 strain is detected by the QIAstat-Dx Respiratory SARS-CoV-2

Panel assay, two signals will be generated and displayed on the QIAstat-Dx Analyzer

1.0 screen: one for Influenza A and a second one for H3 strain.

Note: It is acceptable if only the H3 signal is obtained, which would be indicated as “equivocal”.

 If a pandemic Influenza A/H1N1/2009 strain is detected, two signals will be generated

and displayed on the QIAstat-Dx Analyzer 1.0 screen: one for Influenza A and a second one for H1N1/2009.

Note: It is acceptable if only the H1N1/2009 signal is obtained, which would be indicated as “equivocal”.

Note: It is acceptable if only the Influenza A signal is obtained, which would be indicated as “Influenza A (no subtype detected)”.

IMPORTANT: If only an Influenza A signal is present and no additional signal for any of the subtypes is generated, it can be due to either low concentration or, in very rare cases, a new variant or any Influenza A strain other than H1 and H3 (eg, H5N1, which can infect humans). See important precautions regarding possible detection of Influenza A with no subtype detected.

Result Interpretation for all other pathogens

For every other pathogen that can be detected with the QIAstat-Dx Respiratory SARS-CoV-2

Panel, only one signal will be generated if the pathogen is present in the sample.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Viewing amplification curves

To view test amplification curves of pathogens detected, press the Amplification Curves tab (Figure 17).

Figure 17. Amplification Curves screen (PATHOGENS tab).

Details about the tested pathogens and controls are shown on the left, and the amplification curves are shown in the center.

Note: If User Access Control is enabled on the QIAstat-Dx Analyzer 1.0, the Amplification Curves screen is only available for operators with access rights.

Press the PATHOGENS tab on the left side to display the plots corresponding to the tested pathogens. Press on the pathogen name to select which pathogens are shown in the amplification plot. It is possible to select single, multiple, or no pathogens. Each pathogen in the selected list will be assigned a color corresponding to the amplification curve associated with the pathogen. Unselected pathogens will be shown in gray.

The corresponding C

and endpoint fluorescence (EP) values are shown below each pathogen

name.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Press the CONTROLS tab on the left side to view the controls in the amplification plot. Press the circle next to the control name to select or deselect it (Figure 18).

Figure 18. Amplification Curves screen (CONTROLS tab).

The amplification plot displays the data curve for the selected pathogens or controls. To alternate between logarithmic or linear scale for the Y-axis, press the Lin or Log button at the bottom left corner of the plot.

The scale of the X-axis and Y-axis can be adjusted using the blue pickers on each axis. Press and hold a blue picker and then move it to the desired location on the axis. Move a blue picker to the axis origin to return to the default values.

Viewing test details

Press Test Details in the Tab Menu bar at the bottom of the touchscreen to review the results in more detail. Scroll down to see the complete report. The following Test Details are shown in the center of the screen (Figure 19, next page):

 User ID

 Cartridge SN (serial number)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

 Cartridge Expiration Date

 Module SN (serial number)

 Test Status (Completed, Failed, or Canceled by operator)

 Error Code (if applicable)

 Test Start Date and Time

 Test Execution Time

 Assay Name

 Test ID

 Test Result:

 Positive (if at least one respiratory pathogen is detected/identified)  Positive with warning (at least one respiratory pathogen is detected but the Internal

Control failed)

 Negative (no respiratory pathogen is detected)  Invalid

 List of analytes tested in the assay, with C

and endpoint fluorescence in the event of a

positive signal

 Internal Control, with C

Figure 19. Example screen showing Test Data on the left panel and Test Details in the main panel.

and endpoint fluorescence

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Browsing results from previous tests

To view results from previous tests that are stored in the results repository, press View Results on the Main Menu bar (Figure 20).

Figure 20. Example View Results screen.

The following information is available for every executed test (Figure 21):

 Sample ID

 Assay (name of test assay)

 Operator ID

 Mod (Analytical Module on which the test was executed)

 Date/Time (date and time when the test was finished)

 Result (outcome of the test: positive [pos], positive with warning [pos*], negative [neg],

failed [fail] or successful [suc])

Note: If User Access Control is enabled on the QIAstat-Dx Analyzer 1.0, the data for which the user has no access rights will be hidden with asterisks.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Select one or more test results by pressing the gray circle to left of the sample ID. A checkmark will appear next to selected results. Unselect test results by pressing this checkmark. The entire list of results can be selected by pressing the checkmark circle in the top row (Figure 21).

Figure 21. Example of selecting Test Results in the View Results screen.

Press anywhere in the test row to view the result for a particular test.

Press a column headline (e.g., Sample ID) to sort the list in ascending or descending order according to that parameter. The list can be sorted according to only one column at a time.

The Result column shows the outcome of each test (Table 3):

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 3. Descriptions of test results

Outcome Result Description

Positive

Positive with warning

Negative

Failed

Successful

pos

pos*

neg

fail

suc

At least one pathogen is positive.

At least one pathogen is positive but the Internal Control failed.

No pathogens were detected.

The test failed because either an error occurred or the test was canceled by the user.

The test is either positive or negative, but the user does not have the access rights to view the test results

Make sure a printer is connected to the QIAstat-Dx Analyzer 1.0 and the proper driver is installed. Press Print Report to print the report(s) for the selected result(s).

Press Save Report to save the report(s) for the selected result(s) in PDF format to an external USB storage device.

Select the report type: List of Tests or Test Reports.

Press Search to search the test results by Sample ID, Assay and Operator ID. Enter the search string using the virtual keyboard and press Enter to start the search. Only the records containing the search text will be displayed in the search results.

If the results list has been filtered, the search will only apply to the filtered list.

Press and hold a column headline to apply a filter based on that parameter. For some parameters, such as Sample ID, the virtual keyboard will appear so the search string for the filter can be entered.

For other parameters, such as Assay, a dialog will open with a list of assays stored in the repository. Select one or more assays to filter only the tests that were performed with the selected assays.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

The symbol to the left of a column headline indicates that the column’s filter is active.

A filter can be removed by pressing Remove Filter in the Submenu bar.

Exporting results to a USB drive

From any tab of the View Results screen, select Save Report to export and save a copy of the test results in PDF format to a USB drive. The USB port is located on the front of the QIAstat-Dx Analyzer 1.0.

Printing results

Make sure a printer is connected to the QIAstat-Dx Analyzer 1.0 and the proper driver is installed. Press Print Report to send a copy of the test results to the printer.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Quality Control

In accordance with QIAGEN’s ISO-certified Quality Management System, each lot of QIAstat-Dx Respiratory SARS-CoV-2 Panel is tested against predetermined specifications to ensure consistent product quality. External controls are not provided with the QIAstat-Dx Respiratory SARS-CoV-2 Panel. Quality control requirements should be performed in conformance with local, state, and/or federal regulations or accreditation requirements and your laboratory’s standard quality control procedures.

The following external controls are available:

 ZeptoMetrix

 ZeptoMetrix Inc. NATtrol SARS-CoV-2 E/ORF1ab recombinant (Cat. No. 0831043)

Inc. NATtrol™ Respiratory Verification Panel (Cat. No. NATRVP-QIA)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Limitations

 For prescription use only.

 The use of this assay as an in vitro diagnostic under FDA Emergency Use Authorization

(EUA) is limited to laboratories that are certified under the Clinical Laboratory Improvement Amendments of 1988 (CLIA), 42 U.S.C. §263a, to perform high and moderate complexity tests.

 Results from the QIAstat-Dx Respiratory SARS-CoV-2 Panel are not intended to be used as

the sole basis for diagnosis, treatment, or other patient management decisions.

 Laboratories are required to report all positive SARS-CoV-2 results to the appropriate

public health authorities.

 Primers and probes for this kit target highly conserved regions within the genome of

SARS- CoV-2. Mutations occurring in these highly conserved regions (although rare) may result in RNA being undetectable.

 The E gene target is homologous to sequences from multiple bat SARS viruses. These

viruses are unlikely to be found in nasopharyngeal swabs and their potential to infect human hosts is unknown.

 The performance of this test has not been established for immunocompromised

individuals.

 The performance of this test has not been established for patients without signs and

symptoms of respiratory infection.

 Positive results do not rule out co-infection with organisms not included in the QIAstat-Dx

Respiratory SARS-CoV-2 Panel. The agent detected may not be the definitive cause of the disease.

 Negative results do not preclude infection of the upper respiratory tract. Not all agents of

acute respiratory infection are detected by this assay and sensitivity in some clinical settings may differ from that described in the package insert.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

 A negative result with the QIAstat-Dx Respiratory SARS-CoV-2 Panel does not exclude the

infectious nature of the syndrome. Negative assay results may originate from several factors and their combinations, including sample handling mistakes, variation in the nucleic acid sequences targeted by the assay, infection by organisms not included in the assay, organism levels of included organisms that are below the limit of detection for the assay, and use of certain medications, therapies, or agents.

 The QIAstat-Dx Respiratory SARS-CoV-2 Panel is not intended for testing of samples other

than those described in these Instructions for Use. Test performance characteristics have been established only with nasopharyngeal swab samples collected in universal transport media (UTM) from individuals with acute respiratory symptoms.

 The QIAstat-Dx Respiratory SARS-CoV-2 Panel is intended to be used in conjunction with

standard of care culture for organism recovery, serotyping, and/or antimicrobial susceptibility testing where applicable.

 The results from the QIAstat-Dx Respiratory SARS-CoV-2 Panel must be interpreted by a

trained healthcare professional within the context of all relevant clinical, laboratory, and epidemiological findings.

 The QIAstat-Dx Respiratory SARS-CoV-2 Panel can be used only with the QIAstat-Dx

Analyzer 1.0.

 The QIAstat-Dx Respiratory SARS-CoV-2 Panel is a qualitative assay and does not

provide a quantitative value for detected organisms.

 Viral and bacterial nucleic acids may persist in vivo, even if the organism is not viable or

infectious. Detection of a target marker does not imply that the corresponding organism is the causative agent of the infection or the clinical symptoms.

 Detection of viral and bacterial nucleic acids depends on proper sample collection,

handling, transportation, storage and loading into the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge. Improper operations for any of the aforementioned processes can cause incorrect results, including false-positive or false-negative results.

 The performance of this test has not been established for screening of blood or blood products.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

 The performance of this test has not been established in individuals who received

influenza vaccine. Recent administration of a nasal influenza vaccine may cause false positive results for Influenza A and/or Influenza B.

 The QIAstat-Dx Respiratory SARS-CoV-2 Panel may not be able to distinguish between

existing viral strains and new variants as they emerge. For example, the QIAstat-Dx Respiratory SARS-CoV-2 Panel can detect seasonal H3N2 Influenza but may not be able to distinguish seasonal H3N2 from H3N2 variant (H3N2v).

 The QIAstat-Dx Respiratory SARS-CoV-2 Panel detects the multi-copy IS481 insertion

sequence present in multiple possible if the specimen is contaminated with non-pertussis

 The assay sensitivity and specificity, for the specific organisms and for all organisms

combined, are intrinsic performance parameters of a given assay and do not vary depending on prevalence. In contrast, both the negative and positive predictive values of a test result are dependent on the disease/organism prevalence. False negative test results are more likely during peak activity when prevalence of disease is high. False positive test results are more likely during periods when prevalence is moderate or low.

Bordetella

species. False positive

Bordetella

B. pertussis

species.

results are

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Conditions of Authorization for the Laboratory

The QIAstat-Dx Respiratory SARS-CoV-2 Panel Letter of Authorization, along with the authorized Fact Sheet for Healthcare Providers, the authorized Fact Sheet for Patients, and authorized labeling are available on the FDA website: https://www.fda.gov/MedicalDevices/Safety/EmergencySituations/ucm161496.htm.

However, to assist clinical laboratories using the QIAstat-Dx Respiratory SARS-CoV-2 Panel (“your product” in the conditions below), the relevant Conditions of Authorization are listed below:

 Authorized laboratories * using your product will include with result reports of your

product, all authorized Fact Sheets. Under exigent circumstances, other appropriate methods for disseminating these Fact Sheets may be used, which may include mass media.

 Authorized laboratories using your product will use your product as outlined in the

Instructions for Use. Deviations from the authorized procedures, including the authorized instruments, authorized extraction methods, authorized clinical specimen types, authorized control materials, authorized other ancillary reagents, and authorized materials required to use your product are not permitted.

 Authorized laboratories that receive your product will notify the relevant public health

authorities of their intent to run your product prior to initiating testing.

 Authorized laboratories using your product will have a process in place for reporting test

results to healthcare providers and relevant public health authorities, as appropriate.

* The letter of authorization refers to, “United States (U. S.) laboratories certified under the Clinical Laboratory

Improvement Amendments of 1988 (CLIA), 42 U.S.C. §263a, to perform high and moderate complexity tests” as “authorized laboratories.”

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

 Authorized laboratories will collect information on the performance of your product and

will report to DMD/OHT7-OIR/OPEQ/CDRH (via email: CDRH-EUAReporting@fda.hhs.gov) and QIAGEN (https://www.qiagen.com/us/service-andsupport/technical-support/technical-support-form/) any suspected occurrence of false

positive or false negative results, and significant deviations from the established performance characteristics of your product of which they become aware.

 All laboratory personnel using your product must be appropriately trained in RT-PCR

techniques, use appropriate laboratory and personal protective equipment when handling this kit, and use your product in accordance with the authorized labeling.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Performance Characteristics

The QIAstat-Dx Respiratory SARS-CoV-2 Panel (Cat. No. 691223) assay was developed by introducing the reagents required to detect the SARS-CoV-2 target in a separate reaction chamber of the QIAstat-Dx Respiratory Panel assay (Cat. No. 691221), leaving all other targets unchanged. As a result of this and/or availability of SARS-CoV-2 clinical samples, certain studies shown below were not done or repeated using the QIAstat-Dx Respiratory SARS-CoV-2 Panel.

SARS-CoV-2 Target

Clinical performance

The performance of SARS-CoV-2 target in the QIAstat-Dx Respiratory SARS-CoV-2 Panel was evaluated using retrospective nasopharyngeal swab clinical specimens in transport medium. Specifically, 30 individual negative nasopharyngeal specimens and a total of 30 positive samples consisting of:

 10 positive clinical samples tested with a validated molecular comparator assay

obtained from a Hospital in Barcelona (Spain)

 20 contrived positive clinical samples at 1-2x LoD

All clinical samples were collected from patients with signs and symptoms of upper respiratory

infection by qualified personnel according to the package insert of the collection device and stored frozen until use.

Low positive contrived clinical samples were prepared by spiking a quantified clinical sample obtained from a Hospital in Barcelona (Spain) into individual negative clinical samples to approximately 1x- 2x LoD (20 samples).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Overall results are shown on Table 4:

Table 4. Overall clinical performance results of SARS-CoV-2 target

Sample Sample Type N

Positives Positive clinical sample 10 (10/10) 100% N/A 0/0 N/A

Total Positive Samples 30 (30/30) 100% 85.8 -100 0/0 N/A

Negative Total Negative Samples 30 0/0 N/A (30/30) 100% 85.8 -100

Low positive contrived sample (1x-2xLoD)

% Positive (95% CI) % Negative

(20/20) 100% N/A 0/0 N/A

SARS-CoV-2 Target

(95% CI)

Performance of the SARS-CoV-2 target in the QIAstat-Dx Respiratory SARS-CoV-2 Panel against the expected results are:

Positive Percent Agreement (PPA%): 30/30 = 100 % (95% CI: 85.8% - 100%)

Negative Percent Agreement (NPA%): 30/30 = 100% (95% CI: 85.8% - 100%)

Analytical performance

Sensitivity (Limit of Detection)

The Analytical Sensitivity, or Limit of Detection (LoD), is defined as the lowest concentration at which ≥95% of the tested samples generate a positive call.

The LoD of the SARS-CoV-2 target was assessed by analyzing serial dilutions of analytical samples prepared from a quantified clinical sample obtained from a Hospital from Barcelona (Spain). Dilutions were preformed using simulated matrix consisting of UTM and HeLa cells. Four (4) replicates were tested of each serial dilution. The lowest concentration at which all replicates were positive was interpreted as the tentative LoD. The LoD was then confirmed by testing twenty (20) replicates with concentrations at the tentative limit of detection. To confirm the established LoD concentration, the detection rate of all replicates must be ≥95% (at least 19/20 replicates must generate a positive signal).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 5. LoD value obtained for the SARS-CoV-2 target tested with the QIAstat-Dx Respiratory SARS-CoV-2 Panel

Pathogen Strain Source Concentration Detection rate

SARS-CoV-2 Clinical Sample Hospital from Barcelona (Spain) 500 copies/mL 20/20

The final LoD for the SARS-CoV-2 target in the QIAstat-Dx Respiratory SARS-CoV-2 panel according to the assay results interpretation, is 500 copies/ml.

FDA SARS-CoV-2 reference panel testing

The evaluation of sensitivity and MERS-CoV cross-reactivity was performed using reference material (T1), blinded samples, and a standard protocol provided by the FDA. The study included a range-finding study and a confirmatory study for LoD. Blinded sample testing was used to establish specificity and to confirm the LoD. All the reagents required for the complete execution of the test are pre-loaded and self-contained in the QIAstat-Dx Respiratory SARS-CoV-2 Panel cartridge. The instrument used was the QIAstat-Dx Analyzer 1.0. The results are summarized in Table 6 .

Table 6. Summary of LoD confirmation result using the FDA SARS-CoV-2 Reference Panel

Reference Materials Provided by FDA Specimen Type

SARS-CoV-2

MERS-CoV N/A ND

Abbreviations: NDU/ml = RNA NAAT detectable units/ml; ND = not detected; N/A = not applicable.

Nasopharyngeal swab

Product LoD Cross-Reactivity

1.8x105 NDU/ml N/A

Exclusivity (Cross-reactivity and Exclusivity)

The analytical specificity study was carried out by the cross-reactivity and exclusivity of the SARS-CoV-2 target. On-panel organisms were tested to assess the potential for intra-panel cross-reactivity and off-panel organisms were tested to evaluate panel exclusivity. The off-panel organisms selected were clinically relevant organisms (colonizing the upper respiratory tract or causing respiratory symptoms), common skin flora, or laboratory contaminants, or microorganisms for which much of the population may have been infected.

* Only a limited number of organisms were tested

in vitro

in silico

analysis and

(shown in Table 7).

in vitro *

testing to assess

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Samples were prepared by spiking potential cross-reactive organisms into simulated nasopharyngeal swab sample matrix at the highest concentration possible based on the

organism stock – at least 10

TCID50/ml for viral targets and 106 CFU/ml for bacterial and fungal targets. These concentrations represent levels approximately 800–1,000,000-fold higher than the LoD of the SARS-CoV-2 Target.

Table 7. List of Analytical Specificity pathogens tested

Type Pathogen

On-panel bacteria

Off-panel bacteria

Off-panel viruses

* SARS Coronavirus was tested using custom gBlocks from the two regions targeted by the SARS-CoV-2 designs.

In silico

, sequence hits were analyzed together in order to detect unique specific sequences

in vitro

Chlamydophila pneumoniae

Haemophilus influenzae Streptococcus pyogenes Streptococcus pneumoniae Mycobacterium tuberculosis

MERS Coronavirus SARS Coronavirus*

matching with all primers and probes to be considered as positive amplifications. Primers and probes were considered as reactive if the following parameters were fulfilled:

 At least one forward, one probe, and one reverse primer of the SARS-CoV-2 assay match

with the target BLAST hit sequence.

 At least 70% of query cover/identity between the BLAST hit sequence and every single

primer/probe sequence.

 A maximum of 500 bp of amplicon size.

This analysis of the SARS-CoV-2 designs show that a potential unspecific signal can be produced by a cross-reaction with a group of coronaviruses found in bats. These coronaviruses have only been detected in bats and have not been reported to infect or colonize humans. No unspecific signals were generated with critical off-panel human targets.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

No cross-reactivity was observed, both pathogens (colonizing the upper respiratory tract or causing respiratory symptoms), or common sin flora or laboratory contaminants, or microorganisms.

in silico

and

in vitro

, with any clinically relevant

Inclusivity (Analytical Reactivity)

In silico

analysis shows that the SARS-CoV-2 assays in the QIAstat-Dx Respiratory SARS-CoV2 panel show a 100% sequence identity to 1184 out of the 1196 (99.0%) SARS-CoV-2 genomes available in the public databases.

Twelve genome sequences showed a single nucleotide mismatch with one oligonucleotide sequence of the E gene assay. These single mismatches are tolerated by the PCR workflow in the QIAstat-Dx system and these genetic variants of the SARS-CoV-2 virus are predicted to be detected by the QIAstat-Dx Respiratory SARS-CoV-2 panel despite the mismatches.

Additional Targets included in the QIAstat-Dx SARS-CoV-2 Panel

The performance of the other targets in the panel (Adenovirus, Coronavirus 229E, Coronavirus HKU1, Coronavirus NL63, Coronavirus OC43, Human Metapneumovirus A+B, Influenza A, Influenza A H1, Influenza A H3, Influenza A H1N1/pdm09, Influenza B, Parainfluenza virus 1, Parainfluenza virus 2, Parainfluenza virus 3, Parainfluenza virus 4, Rhinovirus/Enterovirus, Respiratory Syncytial Virus A+B,

Mycoplasma pneumoniae

) has been previously established and is presented below.

Bordetella pertussis, Chlamydophila pneumoniae

, and

Clinical performance

The clinical performance of the QIAstat-Dx Respiratory Panel (Cat. No. 691221) was established during a multi-center study conducted at six (6) geographically diverse study sites: five (5) U.S. sites and one (1) international site. Each study location was representative of the intended use setting (clinical laboratories), and testing was performed by trained clinical laboratory personnel. Residual nasopharyngeal swab (NPS) samples were collected from subjects with signs and symptoms of respiratory infection for QIAstat-Dx Respiratory Panel and comparator testing.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Residual NPS specimens in UTM were tested with the QIAstat-Dx Respiratory Panel (Cat. No. 691221) and an FDA-cleared molecular comparator, in accordance with product instructions for use. Specimens tested in the clinical study were collected using the Universal

™

Transport Medium (UTM) (Copan Diagnostics [Brescia, Italy and CA, USA]), MicroTest

, M5®, M6® (Thermo Fisher Scientific®, MA, USA), BD™ Universal Viral Transport (UVT)

M4RT System (Becton Dickinson, NJ, USA), Universal Transport Medium (UTM) System (HealthLink

Inc., FL, USA), Universal Transport Medium (Diagnostic Hybrids

(Quest Diagnostics

, NJ, USA) and UniTranz-RT® Universal Transport Media (Puritan®

, OH, USA), V-C-M Medium

M4®,

Diagnostics, ME, USA) collection kits.

A total of 2304 residual NPS specimens (1994 prospective, 310 archived) were tested in this comparison study. Between December 2017 to April 2019, specimens were prospectively collected from all comers meeting the study inclusion criteria and immediately frozen for later testing by the study site as frozen prospective specimens (N=1093). No frozen samples were distributed amongst sites. At time of testing, specimens were thawed and tested on both the QIAstat-Dx Respiratory Panel and comparator method.

Between February and August 2018, specimens were prospectively collected from all comers meeting the study eligibility criteria and tested fresh (N=901) on both the QIAstat-Dx Respiratory Panel and comparator method in accordance with product instructions as fresh prospective specimens. One specimen was withdrawn from the study due to an incorrect specimen type.

Table 8 (next page) provides the summary of demographic information for the 1994 subjects that participated in the prospective study.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 8. Demographic summary for the prospective study arm

300

Site 1 Copenhagen, Denmark

186 0 196 177 170 195

232 0 230 271 133 204

126 0 103 49 216 133

110 0 56 107 7 50

148 0 227 254 1 168

272 0 50 44 145 277

145 0 318 0 101 122

Overall

Male 924

(46.3%)

SEX

Female 1070

(53.7%)

≤5 years 627

(31.4%)

6–21 years 239

(11.9%)

AGE

22–49 years 330

(16.5%)

50+ years 798

(40.0%)

Outpatient 788

(39.5%)

Hospitalized 686

(34.4%)

Emergency 67

(3.4%)

ICU 153

(7.7%)

STATUS

Not provided/ unknown

Total

(15.0%)

1994 418 0 426 448 303 399

Site 2 Minneapolis, MN

Site 3 Indianapolis, IN

Site 4 Liverpool, NY

Site 5 Columbus, OH

Site 6 Albuquerque, NM

34 0 40 38 79 48

0 0 9 34 24 0

1 0 49 70 33 0

0 0 0 300 0 0

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

A total of 1994 specimens were evaluated for all panel members in the prospective study. The performance of the QIAstat-Dx Respiratory Panel was evaluated by comparing the QIAstat-Dx Respiratory Panel test results with those from an FDA-cleared multiplexed respiratory pathogen panel.

Positive Percent Agreement (PPA) for each analyte was calculated as 100% x (TP/[TP+FN]). True Positive (TP) indicates that both the QIAstat-Dx Respiratory Panel and the comparator method yielded a “Detected” result of that specific analyte. A False Negative (FN) indicates that the QIAstat-Dx Respiratory Panel was “Not Detected” while the comparator method was “Detected” for the analyte in question. Negative Percent Agreement (NPA) was calculated as 100% x (TN/[TN+FP]). True Negative (TN) indicates that both the QIAstat-Dx Respiratory Panel and the comparator method resulted in “Not Detected” for that specific analyte. A False Positive (FP) indicates that the QIAstat-Dx Respiratory Panel was “Detected” while the comparator method was “Not Detected” for the specific pathogen.

Binomial two-sided 95% Confidence Intervals were calculated using the Wilson Score Method.

The QIAstat-Dx Respiratory Panel prospective performance data in positive percent and negative percent agreements against the comparator methods are presented by analyte in Table 9, next page.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 9. QIAstat-Dx Respiratory Panel prospective clinical performance summary

Analyte TP/(TP+FN) Sensitivity/PPA 95% CI TN/(TN+FP) Specificity/NPA 95% CI

Viruses

Adenovirusa Fresh 55/58 94.8%

Coronavirus 229E

Coronavirus HKU1b

Coronavirus NL63

Coronavirus OC43d

Frozen 31/32 96.9%

Overall 86/90 95.6%

Fresh 8/9 88.9%

Frozen 0/0 N/A N/A 1089/1089 100.0%

Overall 8/9 88.9% 56.5–

Fresh 3/3 100.0% 43.8–

Frozen 48/49 98.0% 89.3–

Overall 51/52 98.1% 89.9–

Fresh 4/5 80.0% 37.6–

Frozen 36/42 85.7% 72.2–

Overall 40/47 85.1% 72.3–

Fresh 3/3 100.0% 43.8–

Frozen 23/26 88.5% 71.0–

Overall 26/29 89.7% 73.6–

85.9–

98.2

84.3–

99.4

89.1–

98.3

56.5–

98.0

100.0

99.6

99.7

96.4

93.3

92.6

100.0

96.0

96.4

833/839 99.3%

1047/1057 99.1%

1880/1896 99.2%

886/886 100.0%

1975/1975 100.0% 99.8–

890/892 99.8% 99.2–

1035/1040 99.5% 98.9–

1925/1932 99.6% 99.3–

890/890 100.0% 99.6–

1046/1048 99.8% 99.3–

1936/1938 99.9% 99.6–

892/892 100.0% 99.6–

1059/1063 99.6% 99.0–

1951/1955 99.8% 99.5–

(continued on next page)

98.4–

99.7

98.3–

99.5

98.6–

99.5

99.6–

100.0

99.6–

100.0

99.9

99.8

100.0

99.9

100.0

99.9

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 9 (continued)

Analyte TP/(TP+FN) Sensitivity/PPA 95% CI TN/(TN+FP) Specificity/NPA 95% CI

Viruses (continued)

Human

Fresh 62/67 92.5% Metapneumoviruse

Frozen 53/55 96.4%

Overall 115/122 94.3%

Rhinovirus/

Enterovirus

Fresh 144/157 91.7%

Frozen 124/137 90.5%

Overall 268/294 91.2%

Influenza Ag Fresh 132/133 99.2%

Frozen 110/111 99.1% 95.1–

Overall 242/244 99.2% 97.0–

Influenza A

Fresh 0/1 0.0% 0.0–

Frozen 0/0 N/A N/A 1089/1089 100.0% 99.6–

Overall 0/1 0.0% 0.0–

Influenza A

Fresh 62/63 98.4% 91.5– H1N1/pdm09j

Frozen 18/18 100.0% 82.4–

Overall 80/81 98.8% 93.3–

83.7–

96.8

87.7–

99.0

88.6–

97.2

86.3–

95.1

84.4–

94.4

87.4–

93.9

95.8–

99.9

99.8

79.3

99.7

100.0

99.8

828/829 99.9%

99.3–

100.0

1030/1034 99.6%

99.0–

99.8

1858/1863 99.7%

99.4–

99.9

715/739 96.8%

95.2–

97.8

941/953 98.7%

97.8–

99.3

1656/1692 97.9%

97.1–

98.5

753/757 99.5%

98.6–

99.8

972/977 99.5% 98.8–

99.8

1725/1734 99.5% 99.0–

99.7

894/894 100.0% 99.6–

100.0

1983/1983 100.0% 99.8–

100.0

826/831 99.4% 98.6–

99.7

1071/1071 100.0% 99.6–

100.0

1897/1902 99.7% 99.4–

99.9

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 9 (continued)

Analyte TP/(TP+FN) Sensitivity/PPA 95% CI TN/(TN+FP) Specificity/NPA 95% CI

Viruses (continued)

Influenza A H3j

Influenza Bk Fresh 64/67 95.5%

Parainfluenza virus 1l

Parainfluenza virus 2

Parainfluenza virus 3m

Fresh 67/67 100.0%

Frozen 89/90 98.9%

Overall 156/157 99.4%

Frozen 58/62 93.5%

Overall 122/129 94.6%

Fresh 3/3 100.0%

Frozen 13/14 92.9% 68.5–

Overall 16/17 94.1% 73.0–

Fresh 2/2 100.0% 34.2–

Frozen 0/0 N/A N/A 1089/1089 100.0% 99.6–

Overall 2/2 100.0% 34.2–

Fresh 102/104 98.1% 93.3–

Frozen 9/9 100.0% 70.1–

Overall 111/113 98.2% 93.8–

94.5–

100.0

82.4–

100.0

93.3–

99.8

87.6–

98.5

84.6–

97.5

89.2–

97.3

43.8–

100.0

98.7

99.0

100.0

99.5

100.0

99.5

825/826 99.9%

992/998 99.4%

1817/1824 99.6%

827/828 99.9%

1026/1026 100.0%

1853/1854 99.9%

892/892 100.0%

1072/1075 99.7% 99.2–

1964/1967 99.8% 99.6–

893/893 100.0% 99.6–

1982/1982 100.0% 99.8–

788/793 99.4% 98.5–

1081/1081 100.0% 99.6–

1869/1874 99.7% 99.4–

(continued on next page)

99.3–

100.0

98.7–

99.7

99.2–

99.8

99.3–

100.0

99.6–

100.0

99.7–

100.0

99.6–

100.0

99.9

100.0

99.7

100.0

99.9

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Chlamydophila

Table 9 (continued)

Analyte TP/(TP+FN) Sensitivity/PPA 95% CI TN/(TN+FP) Specificity/NPA 95% CI

Viruses (continued)

Parainfluenza virus 4n

Respiratory Syncytial Virus

(RSV)

Bacteria

Bordetella pertussis

pneumoniae

Mycoplasma pneumoniae

Fresh 3/3 100.0%

43.8–

892/892 100.0%

100.0

Frozen 0/0 N/A N/A 1087/1089 99.8%

Overall 3/3 100.0%

43.8–

1979/1981 99.9%

100.0

Fresh 73/76 96.1%

88.9–

819/820 99.9%

98.6

Frozen 139/144 96.5%

92.1–

941/945 99.6%

98.5

Overall 212/220 96.4%

93.0–

1760/1765 99.7%

98.1

Fresh 2/2 100.0% 34.2–

Frozen 1/1 100.0% 20.7–

100.0

893/893 100.0% 99.6–

1082/1088 99.4% 98.8–

100.0

Overall 3/3 100.0%

43.8–

1975/1981 99.7%

100.0

Fresh 4/4 100.0%

Frozen 1/1 100.0%

51.0–

100.0

20.7–

891/891 100.0%

1087/1088 99.9%

100.0

Overall 5/5 100.0%

56.6–

1978/1979 99.9%

100.0

Fresh 18/18 100.0%

Frozen 1/1 100.0%

82.4–

100.0

20.7–

875/877 99.8%

1085/1088 99.7%

100.0

Overall 19/19 100.0%

83.2–

1960/1965 99.7%

100.0

99.6–

100.0

99.3–

99.9

99.6–

100.0

99.3–

100.0

98.9–

99.8

99.3–

99.9

100.0

99.7

99.3–

99.9

99.6–

100.0

99.5–

100.0

99.7–

100.0

99.2–

100.0

99.2–

99.9

99.4–

99.9

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Adenovirus was detected in 3/4 FN specimens using an independent molecular method. Adenovirus was detected in

6/16 FP specimens using an independent molecular method.

The single FN specimen was negative for Coronavirus HKU1 when tested using an independent molecular method.

Coronavirus HKU1 was detected 0/7 FP specimens using an independent molecular method.

Coronavirus NL63 was detected in 7/7 FN specimens using an independent molecular method. Coronavirus NL63

was detected in 1/2 FP specimens using an independent molecular method.

The 3 FN specimens were negative for Coronavirus OC43 when tested using an independent molecular method.

Coronavirus OC43 was detected in 3/4 FP specimens using an independent molecular method.

Human metapneumovirus (hMPV) was detected in 4/7 FN specimens using an independent molecular method.

hMPV was detected in 3/5 FP specimens using an independent molecular method.

Rhinovirus was detected in 18/26 FN specimens using an independent molecular method. Rhinovirus was detected

in 14/36 FP specimens using an independent molecular method.

Influenza A was detected in 1/2 FN specimens by an independent molecular method. Three (3) FP samples were not

available for testing. Influenza A was detected in the 3/6 remaining FP samples by an independent molecular method.

Influenza A H1 was detected in 1/1 FN specimen by an independent molecular method. Note: Non-2009 H1 has

not been in circulation since being replaced by the 2009 H1 and thus this discrepancy test result is likely false.

Influenza A H1N1 pdm09 was detected in 1/1 FN by an independent molecular method. Influenza A H1 was

detected in 3/5 FP specimens by an independent molecular method.

Influenza A H3 was detected in 1/1 FN by an independent molecular method. Influenza H3 was detected in 7/7 FP

specimens by an independent molecular method.

Influenza B was detected in 6/6 FN specimens available for testing by an independent molecular method; one

discordant sample was not tested by an independent molecular method. Influenza B was detected in 1/1 FP specimens available for testing by an independent molecular method.

The single FN specimen was negative for Parainfluenza virus 1 by an independent molecular method. Parainfluenza

virus 1 was detected in 3/3 FP specimens by an independent molecular method.

Parainfluenza virus 3 was detected in 1/2 FN specimens by an independent molecular method. Parainfluenza 3 was

detected in 3/5 FP specimens by an independent molecular method.

Parainfluenza virus 4 was detected in 2/2 FP specimens by an independent molecular method.

Respiratory Syncytial Virus was detected in 2/8 FN specimens by an independent molecular method. Respiratory

Syncytial Virus was detected in 3/5 FP specimens by an independent molecular method.

Bordetella pertussis

Chlamydophila pneumoniae

Mycoplasma pneumoniae

was detected in 1/6 FP specimens by an independent molecular method.

was detected in 1/1 FP specimens by an independent molecular method.

was detected in 1/4 specimens by an independent molecular method.

The QIAstat-Dx Respiratory Panel detected a total of 191 specimens with distinctive multiple organism detections (9.6% of all specimens) in the prospective study.

All distinct co-infection combinations, as detected by the QIAstat-Dx Respiratory Panel during the prospective clinical study, are presented in Table 10

(next page).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 10. Distinct co-infection combinations detected by QIAstat-Dx Respiratory Panel in the prospective study

Distinct co-infection combinations detected by the QIAstat-Dx Respiratory Panel

Analyte 1 Analyte 2 Analyte 3 Analyte 4

Adenovirus

Rhinovirus/ Enterovirus

Respiratory Syncytial Virus

Mycoplasma

Coronavirus NL63

12 3

11 1

2 1

2 0 N/A

Total coinfections

pneumoniae

Adenovirus

Coronavirus HKU1

Coronavirus NL63

Coronavirus HKU1

Adenovirus Respiratory

Human Metapneumovirus

Parainfluenza virus 3

Parainfluenza virus 4

Respiratory Syncytial Virus

Rhinovirus/ Enterovirus

Adenovirus

Adenovirus 1 1 Adenovirus (1)

Bordetella

3 1

Syncytial Virus

3 1

Rhinovirus/ Enterovirus

1 1

8 1

Respiratory Syncytial Virus

4 1

Respiratory Syncytial Virus

2 2

1 1 Coronavirus HKU1

1 0 N/A

pertussis

Number of discrepant co-infections

Discrepant analyte(s)

Rhinovirus/Enterovirus (1); Adenovirus (2)

Respiratory Syncytial Virus (1)

Mycoplasma pneumoniae

Coronavirus HKU1 (1)

(1)

Human Metapneumovirus (1)

Coronavirus HKU1, Parainfluenza virus 4 (1)

Coronavirus HKU1 (1)

Rhinovirus/Enterovirus (1)

Bordetella pertussis

(2)

(continued on next page)

(1)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 10 (continued)

Distinct co-infection combinations detected by the QIAstat-Dx Respiratory Panel

Analyte 1 Analyte 2 Analyte 3 Analyte 4

Coronavirus NL63

Coronavirus OC43

Coronavirus 229E

Human Metapneumovirus

Influenza A (no subtype)

Parainfluenza virus 1

Respiratory Syncytial Virus

Rhinovirus/ Enterovirus

Adenovirus

Human Metapneumovirus

Parainfluenza virus 3

Respiratory Syncytial Virus

Rhinovirus/ Enterovirus

Respiratory Syncytial Virus

Adenovirus 2 1 Adenovirus (1)

Respiratory Syncytial Virus

Rhinovirus/ Enterovirus

Respiratory Syncytial Virus

1 0 N/A

2 0 N/A

Rhinovirus/ Enterovirus

4 0 N/A

Respiratory Syncytial Virus

2 2 Rhinovirus/

1 0 N/A

2 0 N/A

9 3 Rhinovirus/

Adenovirus Coronavirus

Adenovirus 1 1 Influenza A,

1 0 N/A

2 0 N/A

229E

Total coinfections

1 1 Adenovirus,

Number of discrepant coinfections

Discrepant analyte(s)

Enterovirus (2)

Enterovirus (3)

Rhinovirus/ Enterovirus (1)

Adenovirus (1)

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 10 (continued)

Distinct co-infection combinations detected by the QIAstat-Dx Respiratory Panel

Analyte 1 Analyte 2 Analyte 3 Analyte 4

Influenza A (no subtype)

Influenza A H1N1/pdm09

Influenza A H3 Adenovirus 2 1 Adenovirus (1)

Influenza A H3 Coronavirus NL63 Parainfluenza

Influenza A H3 Coronavirus NL63

Respiratory Syncytial Virus

Coronavirus NL63 1 0 N/A

Coronavirus OC43

Rhinovirus/ Enterovirus

Respiratory Syncytial Virus

1 0 N/A

Adenovirus 1 1 Adenovirus (1)

2 0 N/A

Bordetella

1 0 N/A

pertussis

1 0 N/A

virus 1

Bordetella

1 0 N/A

1 1

Total coinfections

pertussis

Influenza A H3 Coronavirus NL63 1 1 NL63 (1)

Influenza A H3

Influenza A H3 Coronavirus 229E 1 0 N/A

Influenza B Coronavirus HKU1 3 0 N/A

Influenza B Coronavirus NL63 1 0 N/A

Coronavirus OC43

Rhinovirus/ Enterovirus

Parainfluenza virus 1

Parainfluenza virus 3

Respiratory Syncytial Virus

Adenovirus

4 2

2 0 N/A

1 0 N/A

Respiratory Syncytial Virus

1 1

Number of discrepant co-infections

(continued on next page)

Discrepant analyte(s)

Bordetella pertussis

(1)

Coronavirus OC43, Adenovirus (1)

Rhinovirus/ Enterovirus (2)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Mycoplasma

Table 10 (continued)

Distinct co-infection combinations detected by the QIAstat-Dx Respiratory Panel

Analyte 1 Analyte 2 Analyte 3 Analyte 4

Influenza B Respiratory

Influenza B Rhinovirus/

Mycoplasma pneumoniae

Parainfluenza virus 1

Parainfluenza virus 3

Parainfluenza virus 4

Syncytial Virus

Enterovirus

Coronavirus HKU1

Rhinovirus/ Enterovirus

Adenovirus 1 0 N/A

Respiratory Syncytial Virus

Rhinovirus/ Enterovirus

Adenovirus 3 2 Adenovirus (2)

Adenovirus Rhinovirus/

Human Metapneumovirus

Respiratory Syncytial Virus

Rhinovirus/ Enterovirus

Respiratory Syncytial Virus

Rhinovirus/ Enterovirus

Number of

Total coinfections

2 0 N/A

7 4 Rhinovirus/

1 1 Coronavirus HKU1

1 0 N/A

discrepant coinfections

1 1 Parainfluenza virus 1

2 0 N/A

1 1 Rhinovirus/

pneumoniae

Enterovirus

2 1

14 3

1 0 N/A

2 0 N/A

3 1 Parainfluenza virus 3

(continued on next page)

Discrepant analyte(s)

Enterovirus (4)

(1)

Enterovirus (1)

(1)

Human Metapneumovirus (1)

Parainfluenza virus 3 (1)

Rhinovirus/ Enterovirus (2), Parainfluenza virus 3 (1)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 10 (continued)

Distinct co-infection combinations detected by the QIAstat-Dx Respiratory Panel

Analyte 1 Analyte 2 Analyte 3 Analyte 4

Respiratory Syncytial Virus

Rhinovirus/ Enterovirus

Total co-infections 191 51

Total double infections 166 42

Total triple infections 22 7

Total quadruple infections 3 2

The three organisms most prevalent in multiple detections by the QIAstat-Dx Respiratory Panel

Human Metapneumovirus

Rhinovirus/ Enterovirus

Respiratory Syncytial Virus

Rhinovirus/ Enterovirus

29 6 Rhinovirus/

Adenovirus 2 0 N/A

Adenovirus 1 0 N/A

2 1

Total coinfections

Number of discrepant coinfections

Discrepant analyte(s)

Human Metapneumovirus, Rhinovirus/ Enterovirus (1)

Enterovirus (5), Respiratory Syncytial Virus (1)

in the prospective study were Rhinovirus/Enterovirus (108/191, 56.5%), Respiratory Syncytial Virus (77/191, 40.8%), and Adenovirus (53/191, 27.7%). The prevalence of individual organisms in each multiple detection are shown in Table 11 (next page).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 11. The prevalence of individual organisms in each QIAstat-Dx multiple detection

Analyte Prevalence in multiple detections (N=191)

Viruses

Adenovirus 53 (27.7%)

Coronavirus 229E 3 (1.6%)

Coronavirus HKU1 26 (13.6%)

Coronavirus NL63 16 (8.4%)

Coronavirus OC43 15 (7.9%)

Human Metapneumovirus 24 (12.6%)

Rhinovirus/Enterovirus 108 (56.5%)

Influenza A H1 0 (0.0%)

Influenza A H1N1/pdm09 6 (3.1%)

Influenza A H3 16 (8.4%)

Influenza B 13 (6.8%)

Parainfluenza virus 1 9 (4.7%)

Parainfluenza virus 2 0 (0.0%)

Parainfluenza virus 3 28 (14.7%)

Parainfluenza virus 4 4 (2.1%)

Respiratory Syncytial Virus 78 (40.8%)

Bacteria

Bordetella pertussis

Chlamydophila pneumoniae

Mycoplasma pneumoniae

4 (2.1%)

0 (0.0%)

5 (2.6%)

Additional distinct co-infection combinations detected by the comparator method but not detected by the QIAstat-Dx Respiratory Panel in the prospective clinical trial are presented in Table 12 (next page).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 12. Additional distinct co-infection combinations detected by the comparator method but not by the QIAstat-Dx Respiratory Panel in the prospective study

Distinct co-infection combinations detected by the comparator method

Analyte 1 Analyte 2 Analyte 3

Adenovirus Coronavirus HKU1 Respiratory Syncytial Virus 1

Adenovirus Coronavirus OC43 Coronavirus NL63 1

Adenovirus Respiratory Syncytial Virus Coronavirus NL63 1

Adenovirus Rhinovirus/Enterovirus Respiratory Syncytial Virus 1

Coronavirus HKU1 Coronavirus OC43 1

Coronavirus HKU1 Respiratory Syncytial Virus 1

Coronavirus HKU1 Coronavirus NL63 Respiratory Syncytial Virus 1

Coronavirus HKU1 Coronavirus NL63 1

Coronavirus HKU1 Parainfluenza virus 1 Rhinovirus/Enterovirus 1

Coronavirus NL63 Respiratory Syncytial Virus 1

Coronavirus NL63 Rhinovirus/Enterovirus 1

Coronavirus NL63 Influenza A H3 1

Coronavirus OC43 Respiratory Syncytial Virus 1

Human Metapneumovirus Parainfluenza virus 3 Rhinovirus/Enterovirus 1

Human Metapneumovirus Rhinovirus/Enterovirus 1

Rhinovirus/Enterovirus Adenovirus 1

Rhinovirus/Enterovirus Influenza A H3 2

Rhinovirus/Enterovirus Parainfluenza virus 3 1

Rhinovirus/Enterovirus Parainfluenza virus 3 Respiratory Syncytial Virus 1

Rhinovirus/Enterovirus Parainfluenza virus 4 2

Influenza A H3 Respiratory Syncytial Virus 1

Influenza B Influenza A (Equivocal) 1

Total co-infections 24

Total double infections 16

Total triple infections 8

Total co-infections

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

A total of 1994 prospective clinical specimens were tested and analyzed during the prospective clinical evaluation. Of these, 95.88% (1912/1994) yielded valid results on the first attempt (i.e., first loaded cartridge). Invalid or no result were obtained for the remaining 82 specimens (4.11%). Forty-two (42) specimens were invalid due to cartridge internal control failure (2.11%). Of these, 20 (1.00%) provided a result for positively detected targets and 22 (1.10%) had no detections. For 40 (2.00%) specimens, no results were obtained due to incomplete runs. Of these, 1 specimen was aborted by users (0.05%), 21 were due to instrument errors (1.05%), and 18 were due to cartridge-related errors (0.90%). Seventy-two (72) of the 82 initially failed (no result or invalid); specimens yielded valid results after a single retesting using a new cartridge/sample. The remaining 10 specimens failed on the second attempt (2 due to cartridge failures, 1 due to instrument errors, and 7 due to internal control failures). Of these internal control failures, detected pathogens were reported for 4 specimens.

Preselected archived specimens

Some of the analytes on the QIAstat-Dx Respiratory Panel were of low prevalence and were not encountered in sufficiently large numbers during the prospective study to adequately demonstrate clinical performance. To supplement the results of the prospective clinical study, an evaluation of preselected frozen archived retrospective specimens was performed. The specimens selected for testing had previously tested positive for one of the following targets at the clinical laboratory by their standard of care method:

Bordetella pertussis

229E, Coronavirus HKU1, Coronavirus NL63, Coronavirus OC43, Influenza A H1N1 2009,

Mycoplasma pneumoniae, Chlamydophila pneumoniae

, Parainfluenza virus 1, Parainfluenza virus 2, and Parainfluenza virus 4. Testing was performed by operators who were blinded to the expected test result. A total of 310 clinical samples were included within the frozen archived retrospective sample tested arm. Samples were tested by both the comparator method and QIAstat-Dx Respiratory Panel. If the comparator method did not confirm the preselected target as positive, it was excluded from the data analysis for that target.

, Coronavirus

A summary of the demographic information available for the archived specimens is provided in Table 13 (next page).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 13. Demographic summary for the retrospective study arm

Overall (%)

Male 158 (50.8%)

SEX

Female 152 (49.2%)

≤5 years 139 (44.9%)

6–21 years 85 (27.4%)

22–49 years 53 (17.1%)

AGE

50+ years 33 (10.7%)

Outpatient 224 (72.3%)

Hospitalized 68 (21.9%)

Emergency 8 (2.6%)

ICU 8 (2.6%)

STATUS

Other 2 (0.6%)

Total 310

The QIAstat-Dx Respiratory Panel retrospective specimens testing performance data against the comparator method are provided in Table 14 (next page) by analyte.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 14. Overall retrospective clinical study performance

Analyte TP/(TP+FN) Sensitivity

/PPA

Viruses

95% CI TN/(TN+FP) Specificity

/NPA

95% CI

Adenovirusa 9/9 100.0% 70.1–100.0 297/304 97.8% 95.4–98.9

Coronavirus 229E 26/27 96.3% 81.7–99.3 286/286 100.0% 98.7–100.0

Coronavirus HKU1b 14/14 100.0% 78.5–100.0 298/299 99.7% 98.1–99.9

Coronavirus NL63c 24/24 100.0% 86.2–100.0 286/288 99.3% 97.5–99.8

Coronavirus OC43 28/29 96.6% 82.8–99.4 279/279 100.0% 98.6–100.0

Human Metapneumovirus 2/2 100.0% 34.2–100.0 311/311 100.0% 98.7–100.0

Rhinovirus/

44/49 89.8% 78.2–95.5 254/264 96.2% 93.2–97.9

Enterovirusd

Influenza A 17/17 100.0% 81.5–100.0 296/296 100.0% 98.7–100.0

Influenza A H1 0/0 N/A N/A 313/313 100.0% 98.8–100.0

Influenza A H1N1/pdm09

7/8 87.5% 52.9–97.8 304/304 100.0% 98.8–100.0

Influenza A H3 8/8 100.0% 67.5–100.0 305/305 100.0% 98.8–100.0

Influenza B 1/1 100.0% 20.7–100.0 312/312 100.0% 98.8–100.0

Parainfluenza virus 1 40/40 100.0% 91.2–100.0 267/267 100.0% 98.6–100.0

Parainfluenza virus 2 3/3 100.0% 43.8–100.0 309/309 100.0% 98.8–100.0

Parainfluenza virus 3f 1/4 25.0% 4.6–69.9 309/309 100.0% 98.8–100.0

Parainfluenza virus 4g 22/24 91.7% 74.2–97.7 278/278 100.0% 98.6–100.0

Respiratory Syncytial

11/12 91.7% 64.6–98.5 300/301 99.7% 98.4–99.9

Virus (RSV)h

Bacteria

Bordetella pertussis

Chlamydophila pneumoniae

Mycoplasma

33/33 100.0% 89.6–100.0 261/261 100.0% 98.5–100.0

54/61 88.5% 78.2–94.3 250/250 100.0% 98.5–100.0

25/25 100.0% 86.7–100.0 287/288 99.7% 98.1–99.9

pneumoniae

Adenovirus was detected in 3/5 FP specimens using an independent molecular method. 2 FP did not undergo

discordant analysis.

The single FP Coronavirus HKU1 specimen was negative when tested using an independent molecular method.

The single FP Coronavirus NL63 specimen was negative when tested using an independent molecular method.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Rhinovirus was detected in 1/2 FN when tested using an independent molecular method. Rhinovirus was detected in

4/10 FP specimens using an independent molecular method.

Influenza H1N1 pdm09 was detected in the single FN specimen.

Parainfluenza virus 3 was detected in 1/3 FN specimens by an independent molecular method.

Parainfluenza virus 4 was detected in 1/2 FN specimens by an independent molecular method.

The single FN Respiratory Syncytial Virus was negative for that target by an independent molecular method. The

single FP Respiratory Syncytial Virus was negative for that target by an independent molecular method.

Chlamydophila pneumoniae

was detected in 4/5 FN specimens by an independent molecular method.

Testing of contrived specimen

Influenza A H1, Parainfluenza virus 2, Parainfluenza virus 4, Coronavirus 229E and

Chlamydophila pneumoniae

insufficient to demonstrate system performance. Therefore, contrived specimens were used as surrogate clinical specimens to supplement and test the sensitivity and specificity of the above analytes. Residual negative clinical specimens were spiked with the pathogens at 3x, 5x and 10x LoD levels (50 of each).

Contrived samples were provided a unique study identification number and the individual who contrived the samples did not test them therefore the status of each contrived specimen was unknown at the time of testing. Results of contrived specimen testing are provided in Table 15 (next page).

, despite all prospective and retrospective testing efforts, were

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 15. Contrived specimen results

Positive Predictive Agreement

Analyte

Influenza A H1* 3 24/24 100% 86.2–100

Coronavirus 229E 3 16/16 100% 80.6–100

Parainfluenza virus 2 3 16/16 100% 80.6–100

Parainfluenza virus 4 3 15/16 93.8% 71.7–98.9

Chlamydophila pneumoniae

* One Influenza A H1 strain [VR-897] was initially spiked incorrectly, yielding unexpected results across all LoD

concentrations [3x LoD = 4/8 (50%), 5x LoD = 2/9 (22.2%) and 10x LoD = 6/8 (75.0%). A replacement strain [0810244CFHI] was sent to the testing site for spiking and strain VR-897 was also repeated to confirm that the issue was isolated to a procedural error and not an instrument failure.

x LoD TP/(TP + FN) % 95% CI

5 27/27 100% 87.5–100

10 24/24 100% 86.2–100

5 18/18 100% 82.4–100

10 16/16 100% 80.6–100

5 18/18 100% 82.4–100

10 16/16 100% 80.6–100

5 18/18 100% 82.4–100

10 16/16 100% 80.6–100

3 16/16 100% 80.6–100

5 18/18 100% 82.4–100

10 16/16 100% 80.6–100

Expected values

During the prospective QIAstat-Dx Respiratory Panel (Cat. No. 691221) clinical study, 1994 eligible prospective nasopharyngeal swab (NPS) specimens were collected and tested at five (5) sites across the U.S. (4) and Europe (1) from December 2017 through June 2018. The number and percentage of positive cases, as determined by the QIAstat-Dx Respiratory Panel, calculated by testing site or by age group are presented in Table 16, Table 17, and Table 18 (following pages).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 16. Expected value (EV) (as determined by the QIAstat Dx Respiratory Panel) summary overall and by site for the prospective clinical evaluation (N = number)

Overall

Organism N EV N EV N EV N EV N EV N EV

Viruses

Adenovirus 102 5.1% 44 10.5% 9 2.1% 12 2.7% 30 9.9% 7 1.8%

Coronavirus 229E 8 0.4% 1 0.2% 0 0.0% 0 0.0% 7 2.3% 0 0.0%

Coronavirus HKU1 58 2.9% 4 1.0% 11 2.6% 14 3.1% 12 4.0% 17 4.3%

Coronavirus NL63 42 2.1% 4 1.0% 1 0.2% 15 3.3% 11 3.6% 11 2.8%

Coronavirus OC43 30 1.5% 0 0.0% 5 1.2% 6 1.3% 12 4.0% 7 1.8%

Human Metapneumovirus

Human Rhinovirus/Enterovirus

Influenza A 251 12.6% 120 28.7% 0 0.0% 58 12.9% 38 12.5% 35 8.8%

Influenza A H1 0 0.0% 0 0.0% 0 0.0% 0 0.0% 0 0.0% 0 0.0%

Influenza A H1N1 pdm 2009

Influenza H3 163 8.2% 52 12.4% 0 0.0% 52 11.6% 28 9.2% 31 7.8%

Influenza B 123 6.2% 58 13.9% 0 0.0% 32 7.1% 7 2.3% 26 6.5%

Parainfluenza virus 1 19 1.0% 2 0.5% 1 0.2% 2 0.4% 4 1.3% 10 2.5%

Parainfluenza virus 2 2 0.1% 2 0.5% 0 0.0% 0 0.0% 0 0.0% 0 0.0%

Parainfluenza virus 3 116 5.8% 23 5.5% 19 4.5% 16 3.6% 23 7.6% 35 8.8%

Parainfluenza virus 4 5 0.3% 1 0.2% 0 0.0% 1 0.2% 0 0.0% 3 0.8%

Respiratory Syncytial Virus

Bacteria

Bordetella pertussis

Chlamydophila pneumoniae

Mycoplasma pneumoniae

(n=1994) Site 1 (n=418) Site 2 (n=426) Site 3 (n=448) Site 4 (n=303) Site 5 (n=399)

120 6.0% 42 10.0% 24 5.6% 14 3.1% 14 4.6% 26 6.5%

304 15.2% 59 14.1% 78 18.3% 39 8.7% 53 17.5% 75 18.8%

85 4.3% 67 16.0% 0 0.0% 4 0.9% 10 3.3% 4 1.0%

217 10.9% 64 15.3% 40 9.4% 35 7.8% 40 13.2% 38 9.5%

9 0.5% 2 0.5% 1 0.2% 0 0.0% 6 2.0% 0 0.0%

6 0.3% 2 0.5% 1 0.2% 1 0.2% 1 0.3% 1 0.3%

24 1.2% 19 4.5% 0 0.0% 2 0.4% 1 0.3% 2 0.5%

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 17. Expected value (EV) (as determined by the QIAstat Dx Respiratory Panel) summary by age category for the prospective clinical evaluation (N = number)

Organism Overall (n=1994)

N EV N EV N EV N EV N EV

Viruses

Adenovirus 102 5.1% 78 12.4% 7 2.9% 11 3.3% 6 0.8%

Coronavirus 229E 8 0.4% 4 0.6% 4 1.7% 0 0.0% 0 0.0%

Coronavirus HKU1 58 2.9% 29 4.6% 5 2.1% 8 2.4% 16 2.0%

Coronavirus NL63 42 2.1% 25 4.0% 3 1.3% 5 1.5% 9 1.1%

Coronavirus OC43 30 1.5% 20 3.2% 2 0.8% 4 1.2% 4 0.5%

Human Metapneumovirus

Human Rhinovirus/Enterovirus

Influenza A 251 12.6% 47 7.5% 36 15.1% 64 19.4% 104 13.0%

Influenza A H1 0 0.0% 0 0.0% 0 0.0% 0 0.0% 0 0.0%

Influenza A H1N1 pdm 2009

Influenza H3 163 8.2% 25 4.0% 30 12.6% 35 10.6% 73 9.1%

Influenza B 123 6.2% 11 1.8% 22 9.2% 27 8.2% 63 7.9%

Parainfluenza virus 1 19 1.0% 11 1.8% 0 0.0% 4 1.2% 4 0.5%

Parainfluenza virus 2 2 0.1% 1 0.2% 0 0.0% 0 0.0% 1 0.1%

Parainfluenza virus 3 116 5.8% 70 11.2% 4 1.7% 6 1.8% 36 4.5%

Parainfluenza virus 4 5 0.3% 4 0.6% 0 0.0% 0 0.0% 1 0.1%

Respiratory Syncytial Virus

Bacteria

Bordetella pertussis

Chlamydophila pneumoniae

Mycoplasma pneumoniae

120 6.0% 46 7.3% 3 1.3% 17 5.2% 54 6.8%

304 15.2% 186 29.7% 35 14.6% 22 6.7% 61 7.6%

85 4.3% 20 3.2% 6 2.5% 30 9.1% 29 3.6%

217 10.9% 135 21.5% 11 4.6% 17 5.2% 54 6.8%

9 0.5% 5 0.8% 2 0.8% 0 0.0% 2 0.3%

6 0.3% 1 0.2% 3 1.3% 2 0.6% 0 0.0%

24 1.2% 4 0.6% 6 2.5% 11 3.3% 3 0.4%

≤5 years (n=627)

6–21 years (n=239)

22–49 years (n=330)

>49 years (n=798)

The number and percentage of co-infection cases, as determined by the QIAstat-Dx Respiratory Panel, calculated by age group are presented in Table 17,next page.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 18. Expected value (co-infections as determined by the QIAstat-Dx Respiratory Panel) summary by age group

Number

Co-infection

AdV + HRV/EV + CoV NL63

AdV + HRV/EV 12 (6.28%) 9 2 1 0

AdV + RSV 11 (5.82%) 11 0 0 0

AdV +

pneumoniae

AdV + CoV HKU1

CoV HKU1+ AdV+ RSV

CoV HKU1 + HMPV

CoV HKU1 + PIV 3 + HRV/EV

CoV HKU1 + PIV 4

CoV HKU1 + RSV

CoV HKU1 + HRV/EV + RSV

CoV HKU1 + HRV/EV

CoV NL63 + AdV+ RSV

CoV NL63 + AdV

CoV NL63 +

pertussis

(expected value) overall (n=191)

2 (1.05%) 2 0 0 0

2 (1.05%) 1 1 0 0

3 (1.57%) 3 0 0 0

1 (0.52%) 1 0 0 0

3 (1.57%) 3 0 0 0

1 (0.52%) 1 0 0 0

8 (4.28%) 5 1 1 1

1 (0.52%) 1 0 0 0

4 (2.09%) 2 0 0 2

1 (0.52%) 1 0 0 0

2 (1.05%) 1 1 0 0

<6 years (n=151)

6–21 years (n=12)

22–49 years (n=14)

>49 years (n=14)

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 18 (continued)

Number

Co-infection

CoV NL63 + PIV 1 1 (0.52%) 0 0 1 0

(expected value) overall (n=191)

<6 years (n=151)

6–21 years (n=12)

22–49 years (n=14)

>49 years (n=14)

CoV NL63 + RSV

CoV NL63 + HRV/EV

CoV OC43 + AdV

CoV OC43 + HMPV

CoV OC43 + PIV 3 + HRV/EV

CoV OC43 + RSV

CoV OC43 + HRV/EV + RSV

CoV OC43 + HRV/EV

CoV 229E + RSV

HMPV + AdV 2 (1.05%) 1 0 1 0

HMPV + RSV 2 (1.05%) 1 0 0 1

HMPV + HRV/EV

HMPV + HRV/EV + AdV + CoV 229E

Influenza A (no subtype) + RSV + AdV

Influenza A (no subtype) + RSV

2 (1.05%) 2 0 0 0

1 (0.52%) 1 0 0 0

4 (2.09%) 3 1 0 0

2 (1.05%) 2 0 0 0

2 (1.05%) 1 1 0 0

1 (0.52%) 1 0 0 0

9 (4.71%) 6 0 2 1

1 (0.52%) 1 0 0 0

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 18 (continued)

Co-infection

Influenza A H1N1/pdm09 + CoV NL63

Influenza A H1N1/pdm09 + CoV OC43 + AdV

Influenza A H1N1/pdm09 + HRV/EV

Influenza A H1N1/pdm09 + HRV/EV +

B. pertussis

Influenza A H1N1/pdm09 + RSV

Influenza A H3 + AdV

Influenza A H3 + CoV NL63 + PIV 1

Influenza A H3 + CoV NL63 +

B. pertussis

Influenza A H3 + CoV NL63

Influenza A H3 + CoV OC43 + AdV + RSV

Influenza A H3 + HRV/EV

Influenza A H3 + PIV 1

Influenza A H3 + PIV 3

Number (expected value) overall (n=191)

1 (0.52%) 0 0 1 0

1 (0.52%) 1 0 0 0

2 (1.05%) 1 1 0 0

1 (0.52%) 1 0 0 0

2 (1.05%) 0 0 1 1

1 (0.52%) 1 0 0 0

1 (0.52%) 0 0 0 1

1 (0.52%) 1 0 0 0

4 (2.09%) 2 0 1 1

2 (1.05%) 2 0 0 0

2 (1.05%) 1 0 0 1

<6 years (n=151)

6–21 years (n=12)

22–49 years (n=14)

(continued on next page)

>49 years (n=14)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 18 (continued)

Number

Co-infection

Influenza A H3 + RSV

Influenza A H3 + CoV 229E

Influenza B + CoV HKU1

Influenza B + CoV NL63

Influenza B + RSV

Influenza B + HRV/EV

M. pneumoniae

+ CoV HKU1

M. pneumoniae

+ HRV/EV

PIV 1 + AdV 1 (0.52%) 1 0 0 0

PIV 1 + RSV 1 (0.52%) 1 0 0 0

PIV 1 + HRV/EV 2 (1.05%) 2 0 0 0

PIV 1 + HRV/EV +

pneumoniae

PIV 3 + AdV 3 (1.57%) 3 0 0 0

PIV 3 + AdV + HRV/EV

PIV 3 + HMPV 2 (1.05%) 2 0 0 0

PIV 3 + RSV 2 (1.05%) 2 0 0 0

PIV 3 + HRV/EV 14 (7.33%) 14 0 0 0

PIV 4 + RSV 1 (0.52%) 1 0 0 0

PIV 4 + HRV/EV 2 (1.05%) 2 0 0 0

(expected value) overall (n=191)

1 (0.52%) 0 0 1 0

1 (0.52%) 0 1 0 0

3 (1.57%) 1 0 0 2

1 (0.52%) 0 1 0 0

2 (1.05%) 2 0 0 0

7 (3.67%) 4 1 1 1

1 (0.52%) 0 1 0 0

1 (0.52%) 0 0 1 0

1 (0.52%) 1 0 0 0

3 (1.57%) 3 0 0 0

<6 years (n=151)

6–21 years (n=12)

22–49 years (n=14)

>49 years (n=14)

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 18 (continued)

Number

Co-infection

RSV + HMPV + HRV/EV + AdV

RSV + HMPV + HRV/EV

RSV + HRV/EV 29 (15.18%) 26 0 2 1

HRV/EV + RSV + AdV

(expected value) overall (n=191)

1 (0.52%) 1 0 0 0

2 (1.05%) 1 0 0 1

2 (1.05%) 2 0 0 0

<6 years (n=151)

6–21 years (n=12)

22–49 years (n=14)

>49 years (n=14)

Analytical performance

Sensitivity (Limit of Detection)

The Analytical Sensitivity, or Limit of Detection (LoD), is defined as the lowest concentration at which ≥95% of the tested samples generate a positive call.

The LoD for each QIAstat-Dx Respiratory Panel pathogen was assessed by analyzing serial dilutions of analytical samples prepared from high-titer stocks obtained from commercial

suppliers (ZeptoMetrix and ATCC analytes.

) or artificial samples for commercially unavailable target

The LoD concentration was determined for a total of 51 pathogen strains. The LoD per analyte was determined using selected strains representing individual pathogens that are possible to detect with the QIAstat-Dx Respiratory Panel. To confirm the established LoD concentration, the detection rate of all replicates must be ≥95% (at least 19/20 replicates must generate a positive signal).

At least three different cartridge lots and at least three different QIAstat-Dx Analyzers were used for LoD determination for every pathogen.

Individual LoD values for each QIAstat-Dx Respiratory Panel target is shown in Table 19 (next page).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 19. LoD values obtained for the different respiratory target strains tested with the QIAstat-Dx Respiratory SARS-CoV-2 Panel

Pathogen Strain Source Concentration Detection rate

Influenza A H1N1

Influenza A H3N2 A/Virginia/ATCC6/2012* ATCC VR-1811 0.1 PFU/ml Flu A: 20/20

Influenza A, subtype H1N1/2009/pdm09

Influenza B B/Virginia/ATCC5/2012* ATCC VR-1807 0.03 PFU/ml 20/20

Coronavirus 229E – ATCC VR-740 0.2 TCID50/ml 20/20

Coronavirus OC43 – ATCC VR-1558 0.1 TCID50/ml 20/20

Coronavirus NL63 –

A/New Jersey/8/76 ATCC VR-897 341 CEID

A/Brisbane/59/07

A/New Caledonia/20/99 ZeptoMetrix

A/Wisconsin/67/2005*

A/Port Chalmers/1/73 ATCC VR-810 499.3 CEID50/ml Flu A: 20/20

A/Virginia/ATCC1/2009 ATCC VR-1736 67 PFU/ml Flu A: 20/20

A/SwineNY/03/2009 ZeptoMetrix

B/FL/04/06 ATCC VR-1804 1080 CEID50/ml 20/20

B/Taiwan/2/62 ATCC VR-295 5000 CEID50/ml 19/20

–* ZeptoMetrix

–*

ZeptoMetrix 0810244CFHI

0810036CFHI

ZeptoMetrix 0810252CFHI

0810249CFHI

0810229CFHI

ZeptoMetrix 0810024CFHI

ZeptoMetrix 0810228CFHI

4 TCID50/ml Flu A: 20/20

15 TCID50/ml Flu A: 20/20

3.8 TCID50/ml Flu A: 20/20

56 TCID50/ml Flu A: 20/20

3.6 TCID50/ml 20/20

0.1 TCID50/ml 20/20

0.01 TCID50/ml 20/20

/ml Flu A: 20/20

H1: 20/20

H1: 19/20

H3: 20/20

H1N1: 20/20

Coronavirus HKU1 –* Clinical sample

S510

40,000 copies/ml 20/20

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 18 (continued)

Pathogen Strain Source Concentration Detection rate

Parainfluenza virus 1 (PIV 1)

Parainfluenza virus 2 (PIV 2)

Parainfluenza virus 3 (PIV 3)

Parainfluenza virus 4a (PIV 4a)

Parainfluenza virus 4b (PIV 4b)

Respiratory Syncytial Virus A

Respiratory Syncytial Virus B

Human Metapneumovirus

Adenovirus GB (Adenovirus B3) ATCC VR-3 4993.0 TCID50/ml 20/20

C35* ATCC VR-94 0.2 TCID50/ml 19/20

–

Greer ATCC VR-92 7.3 TCID50/ml 20/20

–* ZeptoMetrix

C 243 ATCC VR-93 2.3 TCID50/ml 20/20

–* ZeptoMetrix

M-25 ATCC VR-1378 0.5 TCID50/ml 20/20

–* ZeptoMetrix

A2* ATCC VR-1540 12.0 PFU/ml 20/20

Long* ATCC VR-26 33.0 PFU/ml 20/20

18537* ATCC VR-1580 0.03 PFU/ml 20/20

CH93(18)-18 ZeptoMetrix

Peru6-2003 (type B2)* ZeptoMetrix

hMPV-16, IA10-2003 (A1) ZeptoMetrix

hMPV-20, IA14-2003 (A2)*

hMPV-3, Peru2-2002 (B1)*

RI-67 (Adenovirus E4)* ATCC VR-1572 15.8 TCID50/ml 20/20

Adenoid 75 (Adenovirus C5)* ATCC VR-5 7331.0 TCID50/ml 20/20

Adenoid 71 (Adenovirus C1)* ATCC VR-1 69.5 TCID50/ml 20/20

Adenoid 6 (Adenovirus C2)* ATCC VR-846 28.1 TCID50/ml 20/20

Tonsil 99 (Adenovirus C6)* ATCC VR-6 88.8 TCID50/ml 20/20

ZeptoMetrix 0810014CFHI

0810015CFHI

0810016CFHI

0810060BCFHI

0810040CFHI

0810159CFHI

0810161CFHI

ZeptoMetrix, 0810163CFHI

ZeptoMetrix, 0810156CFHI

0.2 TCID50/ml 19/20

1.3 TCID50/ml 19/20

11.5 TCID50/ml 20/20

9.5 TCID50/ml 20/20

0.4 TCID50/ml 20/20

0.01 TCID50/ml 19/20

0.5 TCID50/ml 20/20

0.4 TCID50/ml 19/20

1479.9 TCID50/ml 19/20

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 18 (continued)

Pathogen Strain Source Concentration Detection rate

Enterovirus /US/IL/14-18952 (Enterovirus

Rhinovirus 1059 (Rhinovirus B14)* ATCC VR-284 8.9 TCID50/ml 20/20

Mycoplasma pneumoniae

Chlamydia pneumoniae

Bordetella pertussis

* The LoD has been obtained in simulated matrix.

D68)

Echovirus 6* ATCC VR-241 0.9 TCID50/ml 19/20

HGP (Rhinovirus A2) ATCC VR-482 8.9 TCID50/ml 19/20

11757 (Rhinovirus C16)* ATCC VR-283 50.0 TCID50/ml 20/20

Type 1A* ATCC VR-1559 8.9 TCID50/ml 20/20

M129-B7 (type 1)* ATCC 29342 0.1 CCU/ml 20/20

PI 1428 ATCC 29085 1.0 CCU/ml 20/20

TW183 ATCC VR-2282 14.2 IFU/ml 20/20

CWL-029* ATCC VR-1310 120.0 IFU/ml 19/20

I028 ATCC BAA-

18323* ATCC 9797 2.6 CFU/ml 19/20

ATCC VR-1824 8.9 TCID50/ml 19/20

0.3 CFU/ml 20/20

2707

Exclusivity (Cross-reactivity and Exclusivity)

The analytical specificity study was carried out by

in silico

analysis and

in vitro

testing to assess the cross-reactivity and exclusivity of the QIAstat-Dx Respiratory Panel. On-panel organisms were tested to assess the potential for intra-panel cross-reactivity and off-panel organisms were tested to evaluate panel exclusivity. The off-panel organisms selected were clinically relevant organisms (colonizing the upper respiratory tract or causing respiratory symptoms), common skin flora or laboratory contaminants, or microorganisms for which much of the population may have been infected. The on-panel organisms tested are shown in Table 20 (next page).

Samples were prepared by spiking potential cross-reactive organisms into simulated nasopharyngeal swab sample matrix at the highest concentration possible based on the

organism stock – at least 10

TCID50/ml for viral targets and 106 CFU/ml for bacterial and fungal targets. These concentrations represent levels approximately 800–1,000,000-fold higher than the LoD of the QIAstat-Dx Respiratory Panel.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Bordetella pertussis

detection in the QIAstat-Dx Respiratory SARS-CoV-2 Panel targets the IS481 region. As a consequence, a certain level of cross-reactivity with off-panel species and

Bordetella pertussis

observed when high concentrations of

was predicted by preliminary sequence analysis and was

Bordetella holmesii

were tested. In concordance with the CDC guidelines for assays that use the IS481 as a target region, when using QIAstat-Dx Respiratory SARS-CoV-2 Panel if the C

value for

Bordetella pertussis

is CT>29, a confirmatory

specificity test is recommended.

Table 20. List of Analytical Specificity pathogens tested

Type Pathogen

Mycoplasma pneumoniae

On-panel bacteria

Off-panel bacteria

Bordetella pertussis Chlamydia pneumoniae

Acinetobacter calcoaceticus Bordetella avium Bordetella bronchiseptica Bordetella hinzii Bordetella holmesii Bordetella parapertussis Chlamydia trachomatis Corynebacterium diphteriae Enterobacter aerogenes Escherichia coli Haemophilus influenzae Klebsiella oxytoca Klebsiella pneumoniae Lactobacillus acidophilus Lactobacillus plantarum Legionella bozemanii Legionella dumofii Legionella feeleii Legionella longbeacheae Legionella micdadei Legionella pneumophila Moraxella catarrhalis

(O157)

(continued on next page)

Bordetella

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 20 (continued)

Type Pathogen

Mycobacterium tuberculosis* Mycoplasma genitalium Mycoplasma hominis Mycoplasma orale Neisseria elongata Neisseria gonorrhoeae Neisseria meningitidis Proteus mirabilis

Off-panel bacteria (continued)

On-panel viruses

Pseudomonas aeruginosa Serratia marcescens Staphylococcus aureus Staphylococcus epidermidis Stenotrophomonas maltophilia Streptococcus agalactiae Streptococcus pneumoniae Streptococcus pyogenes Streptococcus salivarus Ureaplasma urealyticum

Influenza A H1N1

Influenza A H3N2

Influenza A H1N1/pdm09

Influenza B

Coronavirus 229E

Coronavirus OC43

Coronavirus NL63

Coronavirus HKU1

Parainfluenza virus 1

Parainfluenza virus 2

Parainfluenza virus 3

Parainfluenza virus 4a

RSV A

hMPV A

Adenovirus C

Adenovirus B

†

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 20 (continued)

Type Pathogen

On-panel viruses (continued)

Off-panel viruses

Off-panel fungi

Mycobacterium tuberculosis

†

Coronavirus HKU1 clinical specimen tested.

‡

Bocavirus Type 1 clinical specimens tested.

Middle East Respiratory Syndrome Coronavirus synthetic RNA tested.

genomic DNA tested.

Enterovirus

Rhinovirus

Bocavirus‡

Cytomegalovirus

Epstein-Barr Virus

Herpes Simplex Virus 1

Herpes Simplex Virus 2

Measles Virus

Middle East Respiratory Syndrome Coronavirus§

Mumps

Aspergillus flavus Aspergillus fumigatus Candida albicans Cryptococcus neoformans

Inclusivity (Analytical Reactivity)

Analytical reactivity (Inclusivity) was evaluated with a collection of 127 respiratory pathogen isolates/strains that were selected based on clinical relevance and temporal/geographical diversity. Based on wet testing and Panel primers and probes are specific and inclusive for clinically prevalent and relevant strains

in silico

for each pathogen. Wet testing has been done with the strains listed in Table 20. Every strain has been tested in triplicate with a 100% detection rate for concentrations listed.

Table 21 (following pages) provides details of the respiratory pathogens tested in this study.

analysis, the QIAstat-Dx Respiratory SARS-CoV-2

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 21. List of Analytical Reactivity pathogens tested

Pathogen

Influenza A

Subtype/ serotype

H1N1

H3N2

Strain Source

A/PR/8/34 ATCC VR-1469 3x Influenza A H1

A/New Jersey/8/76* ATCC VR-897 1x Influenza A H1

A/Brisbane/59/07*

A/New Caledonia/20/99*

A/Denver/1/57 ATCC VR-546 0.1x Influenza A H1

A/Weiss/43 ATCC VR-96 0.1x Influenza A H1

A/Fort Monmouth/1/1947 ATCC VR-1754 0.1x Influenza A H1

A/WS/33 ATCC VR-1520 0.1x Influenza A H1

A/Swine/Iowa/15/1930 ATCC VR-333 1x Influenza A H1

A/Mal/302/54 ATCC VR-98 1x Influenza A H1

A/Virginia/ATCC6/2012* ATCC VR-1811 1x Influenza A H3

A/Wisconsin/67/2005* ZeptoMetrix

A/Port Chalmers/1/73* ATCC VR-810 1x Influenza A H3

A/Victoria/3/75 ATCC VR-822 1x Influenza A H3

A/Aichi/2/68 ATCC VR-1680 10x Influenza A H3

A/Hong Kong/8/68 ATCC VR-1679 10x Influenza A H3

A/Alice (recombinant, carries A/England/42/72)

MRC-2 (recombinant A/England/42/72 and A/PR/8/34 strains)

A/Switzerland/ 9715293/2013

A/Wisconsin/15/2009 ATCC VR-1882 1x Influenza A H3

ZeptoMetrix 0810244CFHI

ZeptoMetrix 0810036CFHI

0810252CFHI

ATCC VR-776 10x Influenza A H3

ATCC VR-777 100x Influenza A H3

ATCC VR-1837 1x Influenza A H3

x LoD Detected

1x Influenza A H1

0.3x Influenza A H1

1x Influenza A H3

Result

* Strain tested during LoD verification study. Note: Influenza A/Brisbane/59/07 (ZeptoMetrix, 0810244CFHI) taken as reference strain to calculate the x-fold LoD detected.

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 21 (continued)

Pathogen

Influenza A

Subtype/ serotype

H1N1 (pandemic)

H2N2

H5N3

Strain Source

A/Virginia/ATCC1/2009* ATCC VR-1736 1x Influenza A

A/SwineNY/03/2009* ZeptoMetrix

Canada/6294/09 ZeptoMetrix

Mexico/4108/09 ZeptoMetrix

Netherlands/2629/2009

A/Virginia/ATCC2/2009 ATCC VR-1737 0.1x

A/Virginia/ATCC3/2009 ATCC VR-1738 100x

Swine NY/01/2009

Swine NY/02/2009

A/California/07/2009 NYMC X-179A

Japan/305/1957 (nucleic acid)

Korea/426/1968xPuerto Rico/8/1934 [isolate ID/source=recombinant] (nucleic acid)

A/Duck/Singapore/645/ 1997 [isolate ID/source=avian]

0810249CFHI

0810109CFJHI

0810166CFHI

BEI Resources NR19823

ZeptoMetrix 0810248CFHI

ZeptoMetrix 0810109CFNHI

ATCC VR-1884 0.1x

BEI Resources NR2775

BEI Resources NR9679

BEI Resources NR9682

x LoD Detected

1x Influenza A

3x Influenza A

0.1x Influenza A

0.3x

10x

1x Influenza A (no

0.3x Influenza A (no

Result

H1N1/pdm09

Influenza A H1N1/pdm09

subtype)

Influenza A (no subtype)

* Strain tested during LoD verification study.

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 21 (continued)

Pathogen

Subtype/ serotype

Strain Source

H10N7 Chicken/Germany/N/49

[isolate ID/source=avian]

BEI Resources NR2765

x LoD Detected

Result

10x Influenza A (no

subtype)

(nucleic acid)

Influenza A

H1N2 Recombinant Kilbourne

F63, A/NWS/1934 (HA) x A/Rockefeller

BEI Resources NR9677

100x Influenza A H1

Institute/5/1957 (NA) [isolate ID/source=recombinant] (nucleic acids)

B/Virginia/ATCC5/2012* ATCC VR-1807 1x Influenza B

B/FL/04/06* ATCC VR-1804 1x Influenza B

B/Taiwan/2/62* ATCC VR-295 0.3x Influenza B

B/Allen/45 ATCC VR-102 Not

detected

B/Hong Kong/5/72 ATCC VR-823 Not

detected

†

Negative

B/Maryland/1/59 ATCC VR-296 0.1x Influenza B

Influenza B

Not available

B/GL/1739/54 ATCC VR-103 1x Influenza B

B/Wisconsin/1/2010 ATCC VR-1883 0.1x Influenza B

B/Massachusetts/2/2012 ATCC VR-1813 3x Influenza B

‡

Influenza B or Negative

B/Florida/02/06 ZeptoMetrix

0810037CFHI

Impaired detectability

B/Brisbane/60/2008 BEI Resources NR-

42005

B/Malaysia/2506/2004 BEI Resources NR-

0.1x Influenza B

0.3x Influenza B

9723

* Strain tested during LoD verification study.

†

Both strains are derivative from B/Lee/40 ancestral lineage and, according to

to be detected by the QIAstat-Dx Respiratory Panel.

‡

In silico

analysis showed that this strain should be detected by QIAstat-Dx Respiratory Panel.

in silico

analysis, they were predicted

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 21 (continued)

Pathogen

Coronavirus 229E

Coronavirus OC43

Coronavirus NL63

Coronavirus HKU1

Subtype/ serotype

Not available

Strain Source

Not available* ZeptoMetrix

Not available* ATCC VR-740 0.3x Coronavirus

Not available* ATCC VR-1558 1x Coronavirus

Not available* ZeptoMetrix

Not available*

Not available

Not available*

Not available

Not available QIAGEN

Not available

0810229CFHI

0810024CFHI

ZeptoMetrix 0810228CFHI

BEI Resources NR470

ZeptoMetrix NATRVP-IDI

QIAGEN Barcelona (STATDx) Clinical sample S510†

Barcelona (STATDx) Clinical sample S501†

QIAGEN Barcelona (STATDx) Clinical sample 496†

x LoD Detected

1x Coronavirus

0.3x

1x Coronavirus

Result

229E

OC43

Coronavirus NL63

Coronavirus HKU1

HKU1

Coronavirus HKU1

* Strain tested during LoD verification study.

†

Stock titer not available according to manufacturer.

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 21 (continued)

Pathogen

Parainfluenza virus 1

Subtype/ serotype

Not available

Strain Source

Not available* ZeptoMetrix

0810014CFHI

C35* ATCC VR-94 1x Parainfluenza

Not available ZeptoMetrix

NATRVP-IDI

†

x LoD Detected

1x Parainfluenza

10x Parainfluenza

Result

virus 1

Greer* ATCC VR-92 1x Parainfluenza

virus 2

Parainfluenza virus 2

Parainfluenza virus 3

Not available

Not available*

ZeptoMetrix

0.3x

0810015CFHI

Not available

ZeptoMetrix

0.1x

0810504CFHI

Not available*

ZeptoMetrix

0810016CFHI

C 243* ATCC VR-93 1x

Not available*

ZeptoMetrix NATRVP-IDI

†

0.1x

M-25* ATCC VR-1378 1x

Parainfluenza virus 2

Parainfluenza virus 3

Parainfluenza virus 4

Not available

Not available*

ZeptoMetrix

0.3x

0810060BCFHI

Not available

ZeptoMetrix

0.1x

0810060CFHI

CH 19503 ATCC VR-1377 0.3x

Parainfluenza virus 4

* Strain tested during LoD verification study.

†

Stock titer not available according to manufacturer.

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 21 (continued)

Pathogen

Respiratory Syncytial Virus A+B

Human Metapneumovirus

Subtype/ serotype

Not available

Strain Source

18537* ATCC VR-1580 1x Respiratory Syncytial

A2* ATCC VR-1540 0.3x Respiratory Syncytial

Long* ATCC VR-26 1x Respiratory Syncytial

CH93(18)-18* ZeptoMetrix

Not available

B WV/14617/85 ATCC VR-1400 1x

IA10-2003*

IA14-2003*

Peru2-2002*

Peru6-2003*

IA3-2002

IA27-2004

Peru3-2003

IA18-2003

Peru1-2002 ZeptoMetrix

0810040CFHI

ZeptoMetrix 0810040ACFHI

ZeptoMetrix 0810161CFHI

ZeptoMetrix 0810163CFHI

ZeptoMetrix 0810156CFHI

ZeptoMetrix 0810159CFHI

ZeptoMetrix 0810160CFHI

ZeptoMetrix 0810164CFHI

ZeptoMetrix 0810158CFHI

ZeptoMetrix 0810162CFHI

0810157CFHI

x LoD Detected

1x Respiratory Syncytial

0.1x

10x Human Meta-

Result

virus A+B

Respiratory Syncytial virus A+B

Human Metapneumovirus A+B

pneumovirus A+B

* Strain tested during LoD verification study.

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 21 (continued)

Pathogen

Adenovirus

* Strain tested during LoD verification study.

Subtype/ serotype

Not available

Strain Source

Tonsil 99* ATCC VR-6 1x Adenovirus

GB* ATCC VR-3 0.3x Adenovirus

Adenoid 71* ATCC VR-1 1x Adenovirus

Adenoid 6* ATCC VR-846 0.3x Adenovirus

Adenoid 75* ATCC VR-5 0.3x Adenovirus

RI-67* ATCC VR-1572 0.3x Adenovirus

Huie ATCC VR-863 0.3x Adenovirus

Gomen ATCC VR-7 0.1x Adenovirus

Slobitski ATCC VR-12 10x Adenovirus

AV-1645 [128] ATCC VR-256 0.3x Adenovirus

Compton ATCC VR-716 0.3x Adenovirus

Holden ATCC VR-718 0.3x Adenovirus

Trim ATCC VR-1815 0.3x Adenovirus

Dugan ATCC VR-931 0.1x Adenovirus

Tak (73-3544) ATCC VR-930 3x Adenovirus

x LoD Detected

Result

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 21 (continued)

Pathogen

Enterovirus

Rhinovirus

Subtype/ serotype

Not available

Strain Source

/US/IL/1418952*

D-1 (Cox)* ATCC VR-241 0.3x Rhinovirus/Enterovirus

H ATCC VR-1432 1x Rhinovirus/Enterovirus

M.K. (Kowalik) ATCC VR-168 10x Rhinovirus/Enterovirus

Gregory ATCC VR-41 10x Rhinovirus/Enterovirus

Bastianni ATCC VR-1660 1x Rhinovirus/Enterovirus

Griggs ATCC VR-1311 0.3x Rhinovirus/Enterovirus

Conn-5 ATCC VR-28 0.3x Rhinovirus/Enterovirus

Ohio-1 ATCC VR-29 3x Rhinovirus/Enterovirus

Nancy ATCC VR-30 0.3x Rhinovirus/Enterovirus

CHHE-29 ATCC VR-47 10x Rhinovirus/Enterovirus

Kuykendall [V-024001-012]

1059* ATCC VR-284 1x Rhinovirus/Enterovirus

2060* ATCC VR-1559 0.1x Rhinovirus/Enterovirus

HGP* ATCC VR-482 1x Rhinovirus/Enterovirus

11757* ATCC VR-283 0.3x Rhinovirus/Enterovirus

FEB ATCC VR-483 1x Rhinovirus/Enterovirus

33342 ATCC VR-1663 3x Rhinovirus/Enterovirus

ATCC VR-1824 1x Rhinovirus/Enterovirus

ATCC VR-850 10x Rhinovirus/Enterovirus

x LoD Detected

Result

* Strain tested during LoD verification study.

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 21 (continued)

Pathogen

M. pneumoniae

B. pertussis

C. pneumoniae

* Strain tested during LoD verification study.

†

Stock titer not available according to manufacturer.

Subtype/ serotype

Not available

Interfering substances

Strain Source

PI 1428* ATCC 29085 1x

M129-B7* ATCC 29342 1x

FH strain of Eaton Agent [NCTC 10119]

I028* ATCC BAA-2707 1x

19323* ATCC 9797 1x

10-536 ATCC 10380† 0.3x

TW183* ATCC VR-2282 1x

CWL-029* ATCC VR-1310 1x

AR-39 ATCC 53592 0.3x

ATCC 15531 0.1x

x LoD Detected

Result

Mycoplasma pneumoniae

Bordetella pertussis

Chlamydophila pneumoniae

The effect of potentially interfering substances on the detectability of the QIAstat-Dx Respiratory Panel organisms was evaluated. Thirty (30) potentially interfering substances were added to contrived samples at a level predicted to be above the concentration of the substance likely to be found in an authentic NPS specimen. The contrived samples (also referred to as combined samples) were each comprised of a mix of organisms tested at a concentration of 5x LoD.

Endogenous substances such as whole blood, human genomic DNA and several pathogens were tested alongside exogenous substances like antibiotics, nasal sprays and different workflow contaminants.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

The combined samples were tested with and without addition of an inhibitory substance allowing direct sample-to-sample comparison. Combined samples not spiked with any test substance served as a positive control. Additionally, for substances that may contain genetic material (such as blood, mucin, DNA and microorganisms), negative specimens (blank sNPS sample matrix with no organism mix) were spiked with only the test substance to evaluate the potential for false positive results due to the test substance itself.

Combined samples not spiked with any test substance served as a positive control and blank sNPS sample matrix with no organism mix as negative controls.

All pathogen-containing samples without spiked interferent generated positive signals for all pathogens present in the respective combined sample. Negative signals were obtained for all pathogens not present in the same sample but detected by the QIAstat-Dx Respiratory Panel.

None of the substances tested showed inhibition, except for the nasal influenza vaccines. This was due to the fact that the selection of substances concentration was higher than the concentrations expected to be present in a sample. In addition, nasal influenza vaccines

(Fluenz Tetra and FluMist

) were predicted to be reactive with the QIAstat-Dx Respiratory Panel Influenza A (subtype) and Influenza B assays. Final dilution without observable interfering effect was 0.000001% v/v for both vaccines.

No impact on performance is expected when clinical liquid samples are examined in the presence of the substances tested.

Clinically relevant co-infections testing demonstrated that when at least two QIAstat-Dx Respiratory Panel pathogens of different concentrations are simultaneously present in one sample all targets can be detected by the assay.

The results of interfering substance testing are provided in Table 22 (next page).

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 22. Final highest concentration without observable inhibitory effect

Substance tested Concentration tested Results

Endogenous substances

Human genomic DNA 200 ng/µL 20 ng/µl No Interference

Human blood (+NaCitrate) 1% v/v No Interference

Mucin from bovine submaxillary 1% v/v No Interference

Competitive microorganisms

Staphylococcus aureus

Neisseria meningitidis

Corynebacterium diphtheriae

Human Cytomegalovirus 1.00E+05 TCID50/ml No Interference

Exogenous substances

Tobramycin 0.6 mg/ml No Interference

Mupirocin 2% w/v No Interference

Saline nasal spray with preservatives 1% v/v No Interference

Afrin®, severe congestion nasal spray (Oxymetazoline HCl)

Analgesic ointment (Vicks® VapoRub®) 1% w/v No Interference

Petroleum Jelly (Vaseline®) 1% w/v No Interference

FluMist nasal influenza vaccine 0.00001% v/v Interference

FluMist nasal influenza vaccine 0.000001% v/v No Interference

Fluenz Tetra nasal influenza vaccine 0.00001% v/v Interference

Fluenz Tetra nasal influenza vaccine 0.000001% v/v No Interference

Disinfecting/cleaning substances

Disinfecting wipes ½ inches2/1 ml UTM No Interference

DNAZap™ 1% v/v No Interference

RNaseOUT™ 1% v/v No Interference

Bleach 5% v/v No Interference

Ethanol 5% v/v No Interference

1.00E+06 CFU/ml No Interference

5.00E+04 CFU/ml No Interference

5.00E+03 CFU/ml No Interference

1% v/v No Interference

(continued on next page)

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 22 (continued)

Substance tested Concentration tested Results

Specimen collection materials

Swab Copan 168C 1 swab/1 ml UTM No Interference

Swab Copan FloQ 1 swab/1 ml UTM No Interference

Swab Copan 175KS01 1 swab/1 ml UTM No Interference

Swab Puritan 25-801 A 50 1 swab/1 ml UTM No Interference

VTM Sigma Virocult

VTM Remel® M4RT

VTM Remel M4

VTM Remel M5 100% No Interference

VTM Remel M6 100% No Interference

BD Universal Viral Transport 100% No Interference

100% No Interference

Reproducibility

Reproducibility testing of contrived samples was performed at three test sites including two external sites (LACNY [Laboratory Alliance of Central New York] and INDIANA [Indiana University]) and one internal site (STAT). The study incorporated a range of potential variation introduced by sites, days, replicates, cartridge lots, operators, and QIAstat-Dx analyzers. For each site, testing was performed across 5 days with 4 replicates per day (leading to a total of 20 replicates per target, concentration and site), a minimum of 2 different QIAstat-Dx Analyzers per site, and at least 2 operators on each testing day.

A total of 12 sample mixes were prepared with at least 3 replicates tested per sample mix. Each pathogen was spiked into HeLa in UTM combined samples in a final concentration of

0.1x LoD, 1x LoD or 3x LoD, respectively. A summary of results for each analyte is provided in Table 23 (next page).

Table 23 summarizes the results for 0.1x LoD concentration where it is observed that the detection rate for 24 of the 24 targets was <95% and therefore the acceptance criteria is met.

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Reproducibility and repeatability will impact the SARS-CoV-2 target in the same manner as other target organisms verified in the QIAstat-Dx Respiratory Panel.

Table 23. Detection rate per target at 0.1x LoD concentration for each site of reproducibility study and 2-sided 95% Confidence Interval by target

Target (0.1x LoD) Site

STAT 10/20 50.0% 29.9–70.1%

Adenovirus (ATCC VR-3)

B. pertussis

(BAA-2707)

C. pneumoniae

(ATCC VR-

2282)

Coronavirus 229E (ATCC VR-

740)

Coronavirus HKU1 (NATRVPIDI)

Coronavirus NL63 (0810228CFHI)

LACNY 9/19 47.4% 27.3–68.3%

INDIANA 10/19 52.6% 31.7–72.7%

All sites (overall) 29/58 50.0% 37.5–62.5%

STAT 9/20 45.0% 25.8–65.8%

LACNY 7/19 36.8% 19.1–59.0%

INDIANA 9/20 45.0% 25.8–65.8%

All sites (overall) 25/59 42.4% 30.6–55.1%

STAT 11/20 55.0% 34.2–74.2%

LACNY 11/19 57.9% 36.3–76.9%

INDIANA 14/20 70.0% 48.1–85.5%

All sites (overall) 36/59 61.0% 48.3–72.4%

STAT 9/20 45.0% 25.8–65.8%

LACNY 12/19 63.2% 41.0–80.9%

INDIANA 5/20 25.0% 11.2–46.9%

All sites (overall) 26/59 44.1% 32.2–56.7%

STAT 17/20 85.0% 64.0–94.8%

LACNY 10/19 52.6% 31.7–72.7%

INDIANA 9/20 45.0% 25.8–65.8%

All sites (overall) 36/59 61.0% 48.3–72.4%

STAT 13/20 65.0% 43.3–81.9%

LACNY 12/19 63.2% 41.0–80.9%

INDIANA 14/19 73.7% 51.2–88.2%

All sites (overall) 39/58 67.2% 54.4–77.9%

Detection rate (# positive)

% detection rate (# positive)

95% Confidence Interval

(continued on next page)

100

QIAstat-Dx Respiratory SARS-CoV-2 Panel Instructions for Use (Handbook) 09/2020

Table 23 (continued)

Target (0.1x LoD)

Coronavirus OC43 (ATCC VR-1558)

Enterovirus (ATCC VR-1824)

Human Metapneumovirus (0810161CF)

Influenza A (0810249CFHI)

Influenza A (ATCC VR-810)

Influenza A (ATCC VR-897)

Site

STAT 13/20 65.0% 43.3–81.9%

LACNY 15/20 75.0%

INDIANA 15/20 75.0% 53.1–88.8%

All sites (overall) 43/60 71.7% 59.2–81.5%

STAT 8/20 40.0% 21.9–61.3%

LACNY 6/19 31.6% 15.4–54.0%

INDIANA 7/20 35.0% 18.1–56.7%

All sites (overall) 21/59 35.6% 24.6–48.3%

STAT 6/20 30.0% 14.5–51.9%

LACNY 9/19 47.4% 27.3–68.3%

INDIANA 9/20 45.0% 25.8–65.8%

All sites (overall) 24/59 40.7% 29.1–53.4%

STAT 19/20 95.0% 76.4–99.1%

LACNY 18/20 90.0% 69.9–97.2%

INDIANA 20/20 100% 83.9–100%

All sites (overall) 57/60 95.0% 86.3–98.3%

STAT 10/20 50.0% 29.9–70.1%

LACNY 9/19 47.4% 27.3–68.3%

INDIANA 16/19 84.2% 62.4–94.5%

All sites (overall) 35/58 60.3% 47.5–71.9%

STAT 14/20 70.0% 48.1–85.5%

LACNY 9/19 47.4% 27.3–68.3%

INDIANA 12/20 60.0% 38.7–78.1%

All sites (overall) 35/59 59.3% 46.6–70.9%

Detection rate (# positive)

% detection rate (# positive)

95% Confidence Interval

53.1–88.8%

(continued on next page)

QIAGEN QIAstat-Dx User Manual

Specifications and Main Features

Frequently Asked Questions

User Manual

Intended Use

Summary and Explanation

QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge Description

Pathogen Information

Principle of the Procedure

Description of the process

Sample collection and cartridge loading

Sample preparation, nucleic acid amplification and detection

Materials Provided

Kit contents

Materials Required but Not Provided

Warnings and Precautions

Safety information

Reagent Storage and Handling

Specimen Handling, Storage and Preparation

Procedure

Internal Control

Protocol: Transport medium liquid samples

Sample collection, transport and storage

Loading a sample into the QIAstat-Dx Respiratory SARS-CoV-2 Panel Cartridge

Starting the QIAstat-Dx Analyzer 1.0

Running a test

Viewing results

Interpretation of Results

Internal Control interpretation

Pathogen Result interpretation

Result interpretation information for SARS-CoV-2:

Result Interpretation information for Influenza A

Result Interpretation for all other pathogens

Viewing amplification curves

Viewing test details

Browsing results from previous tests

Exporting results to a USB drive

Printing results

Quality Control

Limitations

Conditions of Authorization for the Laboratory

Performance Characteristics

SARS-CoV-2 Target

Clinical performance

Analytical performance

Sensitivity (Limit of Detection)

FDA SARS-CoV-2 reference panel testing

Exclusivity (Cross-reactivity and Exclusivity)

Inclusivity (Analytical Reactivity)

Additional Targets included in the QIAstat-Dx SARS-CoV-2 Panel

Clinical performance

Preselected archived specimens

Testing of contrived specimen

Expected values

Analytical performance

Sensitivity (Limit of Detection)

Exclusivity (Cross-reactivity and Exclusivity)

Inclusivity (Analytical Reactivity)

Interfering substances

Reproducibility