DiagCORE® Respiratory Panel 2
Instructions for use
PL-013-05/2018-EN
210002
2 DiagCORE Respiratory Panel 2 Instructions for Use
DiagCORE® Respiratory Panel 2 Instructions for Use (CE)
Cat # RP1: 210002
2018 May
This document is to be used solely for the purpose of analyzing a DiagCORE® Respiratory Panel 2
Cartridge on the DiagCORE® Analyzer. No part of this document may be reproduced or
transmitted by any means, whether electronic or mechanical, or for any purpose, without the
express written permission of STAT-Dx Life, S.L.
© Copyright 2018, STAT-Dx Life, S.L. All rights reserved.
A printed version of these instructions for use is available upon request.
This Instructions for Use can be accessed at http.support.qiagen.com.
STAT-Dx Life, S.L.
Baldiri Reixac 4
08028 Barcelona, Spain
Customer and technical support
If assistance is required, contact Technical Support using the contact information below.
Website: http.support.qiagen.com
When contacting Technical Support about an error with the DiagCORE® Analyzer or about an error
with the DiagCORE® Respiratory Panel 2 Cartridge, note the steps leading up to the error and any
information appearing in any dialog boxes. This will help us solve the problem.
When calling Technical Support about errors, please have the following information ready:
Analyzer serial number, type and version (if applicable)
Error code (if applicable)
Time point when the error occurred for the first time
Frequency of error occurrence (i.e., intermittent or persistent error)
Photo of error, if possible
Copy of support package
DiagCORE Respiratory Panel 2 Instructions for Use 3
Contents
Customer and technical support ............................................................................. 2
Contents ...................................................................................................................... 3
Table of symbols ......................................................................................................... 5
1 Information about the DiagCORE® Respiratory Panel 2 ............................. 6
1.1 Product name and intended use .................................................................... 6
1.2 Package contents .............................................................................................. 7
1.3 Storage and expiration date ............................................................................ 7
2 Safety information ......................................................................................... 8
3 General background .................................................................................... 9
3.1 DiagCORE® Respiratory Panel 2 Cartridge description ................................ 9
3.2 Detected pathogens ....................................................................................... 10
3.3 Description of the procedure ......................................................................... 12
3.4 Sample collection and cartridge loading .................................................... 13
3.5 Sample preparation, nucleic acid amplification, and detection ............ 13
4 Required materials and equipment that are not provided ..................... 15
5 Procedure for dry swab samples ............................................................... 16
5.1 Sample collection, transport, and storage ................................................... 16
5.2 Loading a sample into the DiagCORE® Respiratory Panel 2 Cartridge ... 16
5.3 Starting the DiagCORE® Analyzer .................................................................. 19
5.4 Running a test ................................................................................................... 19
6 Procedure for transport medium liquid samples ...................................... 25
6.1 Sample collection, transport, and storage ................................................... 25
6.2 Loading a sample into the DiagCORE® Respiratory Panel 2 Cartridge ... 25
6.3 Starting the DiagCORE® Analyzer .................................................................. 28
6.4 Running a test ................................................................................................... 29
7 Internal control ............................................................................................ 34
8 Interpretation of results ............................................................................... 35
8.1 Viewing results ................................................................................................... 35
8.2 Result interpretation ......................................................................................... 40
8.3 Internal Control interpretation ........................................................................ 41
4 DiagCORE Respiratory Panel 2 Instructions for Use
9 Installing the Assay Definition File .............................................................. 42
10 Limitations of the procedure .................................................................... 44
11 Performance characteristics. Clinical Performance ............................. 46
12 Performance characteristics. Analytical Performance ......................... 51
12.1 Sensitivity (Limit of Detection) ......................................................................... 51
12.2 Assay Robustness .............................................................................................. 52
12.3 Exclusivity (Analytical Specificity) ................................................................... 52
12.4 Inclusivity (Analytical Reactivity) .................................................................... 54
12.5 Co-Infections ..................................................................................................... 57
12.6 Interfering Substances ..................................................................................... 58
12.7 Carryover ........................................................................................................... 59
12.8 Reproducibility .................................................................................................. 59
12.9 Sample Stability ................................................................................................ 62
13 References ................................................................................................. 63
14 Appendices ............................................................................................... 64
14.1 Disposal information ......................................................................................... 64
14.2 Declaration of conformity ............................................................................... 64
14.3 Glossary .............................................................................................................. 64
14.4 Trademarks and copyright .............................................................................. 65
14.5 Copyright, disclaimer, and warranty ............................................................. 65
14.6 Patent statement ............................................................................................. 65
14.7 Software license agreement .......................................................................... 65
DiagCORE Respiratory Panel 2 Instructions for Use 5
Table of symbols
The following symbols can be found on the DiagCORE® Respiratory Panel 2 Cartridge, packaging,
and these instructions for use.
Description
Legal manufacturer
In vitro diagnostic medical device
CE marking for European Conformity
Catalog number
Storage temperature limitations
Consult instructions for use
Health hazard/hazardous to the ozone layer
Expiration date (YYYY MM DD)
Batch code
Serial number
Do not reuse
Keep away from sunlight
Sufficient for <n> tests
Do not use if package is damaged
Corrosive
n
6 DiagCORE Respiratory Panel 2 Instructions for Use
1 Information about the DiagCORE
®
Respiratory
Panel 2
1.1 Product name and intended use
The DiagCORE® Respiratory Panel 2 is a qualitative test intended for analyzing nasopharyngeal
swab (NPS) samples taken from patients suspected of respiratory infection for the presence of viral
or bacterial nucleic acids. The DiagCORE® Respiratory Panel 2 is able to accept both dry swabs
and transport medium liquid samples. The assay is designed for use with the DiagCORE® Analyzer
for integrated nucleic acid extraction and multiplex, RT-real-time PCR detection.
The following pathogens can be detected and differentiated with the DiagCORE® Respiratory
Panel 2: Influenza A, Influenza A subtype H1N1/2009, Influenza A subtype H1, Influenza A subtype
H3, Influenza B, Coronavirus 229E, Coronavirus HKU1, Coronavirus NL63, Coronavirus OC43,
Parainfluenza virus 1, Parainfluenza virus 2, Parainfluenza virus 3, Parainfluenza virus 4, Respiratory
Syncytial virus A/B, human Metapneumovirus A/B, Adenovirus, Bocavirus, Rhinovirus/Enterovirus,
Mycoplasma pneumoniae, Legionella pneumophila and Bordetella pertussis.
The results from the DiagCORE® Respiratory Panel 2 must be interpreted within the context of all
relevant clinical and laboratory findings.
Assay performance characteristics have been established only for individuals who have shown
respiratory symptoms.
The DiagCORE® Respiratory Panel 2 is intended for professional use only and is not intended for self-
testing.
For in vitro diagnostic use.
DiagCORE Respiratory Panel 2 Instructions for Use 7
1.2 Package contents
DiagCORE® Respiratory Panel 2
Catalog No.
Number of tests
210002
6
DiagCORE® Respiratory Panel 2 cartridge
6 individually packaged cartridges containing
all reagents needed for sample preparation
and multiplex RT-real time PCR plus internal
control
Cartridge composition:
The DiagCORE® Respiratory Panel 2 Cartridge contents includes the following: Ethanol, isopropanol,
enzymes, oligonucleotides, tris-EDTA, sodium azide, guanidine hydrochloride, guanidinium
thiocyanate, guanidine chloride, propylene glycol, triton-x-100 and D-(+)-trehalose dihydrate.
Transfer pipettes
6 individually packaged transfer pipettes for
dispensing liquid sample into the DiagCORE®
Respiratory Panel 2 Cartridge
1.3 Storage and expiration date
Store the DiagCORE® Respiratory Panel 2 Cartridges in a dry, clean storage space at room
temperature (15–25°C). Do not remove the DiagCORE® Respiratory Panel 2 Cartridges or the
transfer pipettes from their individual packaging until actual use. Under these conditions,
DiagCORE® cartridges can be stored until the expiration date printed on the individual packaging.
The expiration date is also included in the cartridge barcode and is read by the DiagCORE®
Analyzer when the cartridge is inserted into the instrument to run a test.
8 DiagCORE Respiratory Panel 2 Instructions for Use
2 Safety information
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.
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 as the
Clinical and Laboratory Standards Institute (CLSI) Protection of Laboratory Workers from
Occupationally Acquired Infections, Approved Guidelines M29, or other appropriate documents
provided by:
OSHA: Occupational Safety and Health Administration (United States of America)
ACGIH: 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,
DiagCORE® cartridges, and transfer pipettes according to the appropriate regulations.
The DiagCORE® cartridge is a closed, single-use device that contains all reagents needed for
sample preparation and multiplex RT- real time PCR within the DiagCORE® Analyzer. Do not use a
DiagCORE® 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 like the Biosafety in Microbiological and Biomedical
Laboratories from the Centers for Disease Control and Prevention and the National Institutes of
Health (www.cdc.gov/od/ohs/biosfty/biosfty.htm).
DiagCORE Respiratory Panel 2 Instructions for Use 9
3 General background
3.1 DiagCORE
®
Respiratory Panel 2 Cartridge description
The DiagCORE® Respiratory Panel 2 Cartridge is a disposable plastic device that allows
performance of fully automated molecular assays for the detection of respiratory pathogens. Main
features of the DiagCORE® Respiratory Panel 2 Cartridge include compatibility with the respiratory
dry swabs (Copan FLOQSwabs, Ref. 503CS01) and 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 DiagCORE® Respiratory Panel 2 Cartridge. The user does not need to come in contact with
and/or manipulate any reagents. During the test, reagents are handled in the Analytical Module
of the DiagCORE® Analyzer by pneumatically-operated microfluidics and make no direct contact
with the Analyzer actuators. The Analyzer 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.
After the DiagCORE® Respiratory Panel 2 Cartridge containing the sample is introduced into the
Analyzer, 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 RT-real-time, multiplex PCR testing within each reaction chamber. An increase
in fluorescence, indicating detection of the target analyte, is detected directly within each
reaction chamber.
10 DiagCORE Respiratory Panel 2 Instructions for Use
The layout of the DiagCORE® Respiratory Panel 2 Cartridge and its features are illustrated below.
DiagCORE® Respiratory Panel 2 Cartridge.
3.2 Detected pathogens
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 DiagCORE® Respiratory Panel 2 Cartridge is a single-use cartridge that includes all reagents
needed for nucleic acid extraction, nucleic acid amplification, detection, and identification of 22
viral and bacterial pathogens that cause respiratory symptoms. Testing requires a small sample
volume and minimal hands-on time, and the results are available in approximately one hour.
Pathogens that can be detected and identified with the DiagCORE® Respiratory Panel 2 are listed
in Table 1.
DiagCORE Respiratory Panel 2 Instructions for Use 11
Table 1. Pathogens detected by the DiagCORE® Respiratory Panel 2
Pathogen
Classification (genome type)
Influenza A
Orthomyxovirus (RNA)
Influenza A, subtype H1N1/2009
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)
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)
Bocavirus
Parvovirus (DNA)
Rhinovirus/Enterovirus
Picornavirus (RNA)
Mycoplasma pneumoniae
Bacterium (DNA)
Legionella pneumophila
Bacterium (DNA)
Bordetella pertussis
Bacterium (DNA)
Note: Enterovirus and rhinovirus are both detected but not differentiated in the DiagCORE®
Respiratory Panel 2.
12 DiagCORE Respiratory Panel 2 Instructions for Use
3.3 Description of the procedure
Diagnostic tests with the DiagCORE® Respiratory Panel 2 are performed on the DiagCORE®
Analyzer. Samples are collected and loaded manually into the DiagCORE® cartridge:
Option 1: Inserting the swab into the swab port in the case of dry swab sample type.
Option 2: Inserting the liquid into the main port using a transfer pipette in the case of
transport medium liquid sample type.
All of the sample preparation and analysis steps are performed automatically by the DiagCORE®
Analyzer.
DiagCORE Respiratory Panel 2 Instructions for Use 13
3.4 Sample collection and cartridge loading
The collection of samples and their subsequent loading into the DiagCORE® 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 only in the case of transport medium
liquid sample type.
3. The sample information is manually written on or a sample label is affixed to the top of a
DiagCORE® cartridge.
4. Sample is loaded manually into the DiagCORE
®
cartridge:
Dry swab sample type: The nasopharyngeal swab sample is inserted into the swab port of
the DiagCORE® cartridge.
Transport medium liquid sample type: 300 µL of the sample is transferred into the
DiagCORE® cartridge using a transfer pipette.
In the case of transport medium liquid sample loading, the user performs a visual check of
the sample inspection window (see image below) to confirm that the liquid sample has
been loaded.
5. The sample barcode and DiagCORE
®
cartridge QR code are scanned in the DiagCORE®
Analyzer.
6. The DiagCORE
®
cartridge is introduced into the DiagCORE® Analyzer.
7. The test is started on the DiagCORE
®
Analyzer.
3.5 Sample preparation, nucleic acid amplification, and detection
The extraction, amplification, and detection of nucleic acids in the sample are performed
automatically by the DiagCORE® Analyzer.
1. The liquid sample is homogenized, and cells are lysed in the lysis chamber of the DiagCORE
®
cartridge, which includes a rotor that turns at high speed.
14 DiagCORE Respiratory Panel 2 Instructions for Use
2. Nucleic acids are purified from the lysed sample via binding to a silica membrane in the
purification chamber of the DiagCORE® 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 DiagCORE®
cartridge.
4. The mixture of sample and PCR reagents is dispensed into the DiagCORE
®
cartridge PCR
chambers, which contain lyophilized, assay-specific primers and probes.
5. The DiagCORE
®
Analyzer creates the optimal temperature profiles to carry out effective RT-real
time PCR and performs real-time fluorescence measurements to generate amplification curves.
6. The DiagCORE
®
Analyzer Software interprets the resulting data and process controls and
delivers a test report.
DiagCORE Respiratory Panel 2 Instructions for Use 15
4 Required materials and equipment that are
not provided
The DiagCORE® Respiratory Panel 2 is designed for use with the DiagCORE® Analyzer. Before
beginning a test, make sure the following are available:
DiagCORE
®
Analyzer (at least one Operational module and one Analytical module)
DiagCORE
®
Analyzer User Manual
DiagCORE
®
Latest Assay Definition File software for Respiratory Panel 2 installed in the
Operational module
16 DiagCORE Respiratory Panel 2 Instructions for Use
5 Procedure for dry swab samples
5.1 Sample collection, transport, and storage
1. Collect nasopharyngeal swab samples according to the swab manufacturer’s recommended
procedures.
5.2 Loading a sample into the DiagCORE
®
Respiratory Panel 2 Cartridge
1. Open the package of a DiagCORE
®
Respiratory Panel 2 Cartridge using the tear notches on
the sides of the packaging.
After the package is open, sample should be introduced inside the DiagCORE® Respiratory
Panel 2 cartridge and loaded in the DiagCORE® Analyser within 120 minutes.
2. Take out the cartridge and position it so that the QR code on the label faces you.
DiagCORE Respiratory Panel 2 Instructions for Use 17
3. Manually write the sample information or place a sample information label on the top of the
DiagCORE® cartridge. Make sure that the label is properly positioned and does not block the
lid opening.
4. Open the sample lid of the swab port on the left side of the cartridge.
5. Insert the swab into the cartridge until the breakpoint is aligned with the access opening (the
swab will go no further).
18 DiagCORE Respiratory Panel 2 Instructions for Use
6. Break the swab shaft at the breakpoint, leaving the rest of the swab in the cartridge.
7. Firmly close the lid of the swab port until it clicks.
DiagCORE Respiratory Panel 2 Instructions for Use 19
After sample is introduced inside the DiagCORE® Respiratory Panel 2 cartridge, cartridge must
be loaded in the DiagCORE® Analyser within 90 minutes.
5.3 Starting the DiagCORE
®
Analyzer
1. Press the On/Off button on the front of the Analyzer to start the unit.
NOTE
The power switch at the back of the Analytical Module must be set in the “I”
position. The DiagCORE® Analyzer status indicators turn blue in this case.
2. Wait until the Main screen appears and the DiagCORE
®
Analyzer status indicators turn green
and stop blinking.
3. Log in to the DiagCORE
®
Analyzer 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.
4. In case the Assay Definition File software has not been installed in the Analyzer follow the
instructions prior to running the test (See Section 9 of this document for additional information)
5.4 Running a test
1. Press the Run Test button at the top right corner of the touchscreen of the DiagCORE
®
Analyzer.
2. When prompted, scan the sample ID barcode on the nasopharyngeal swab, or scan the
specimen information barcode placed on the top of the DiagCORE® cartridge, using the front
barcode reader that is integrated into the DiagCORE® Analyzer See figure below).
20 DiagCORE Respiratory Panel 2 Instructions for Use
NOTE
It is also possible to enter the sample ID using the virtual keyboard of the
touchscreen by pressing on 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 DiagCORE® Analyzer appear in the Instructions Bar at
the bottom of the touchscreen.
3. When prompted, scan the barcode of the DiagCORE
®
Respiratory Panel 2 Cartridge to be used.
The DiagCORE® Analyzer automatically recognizes the assay to be run, based on the cartridge
barcode.
DiagCORE Respiratory Panel 2 Instructions for Use 21
NOTE
The DiagCORE® Analyzer will not accept cartridges with elapsed expiration
date, 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
cartridge will be rejected. Refer to the DiagCORE® Analyzer User Manual for
further details on how to install assays.
4. Select the appropriate sample type from the list (See image below).
22 DiagCORE Respiratory Panel 2 Instructions for Use
5. The Confirm screen will appear. Review the data entered and make any necessary changes
by pressing the relevant fields on the touchscreen and editing the information.
6. Press Confirm when all the displayed data are correct. If needed, press on the appropriate field
to edit its content, or press Cancel to cancel the test.
7. Ensure that both sample lids of the cartridge are firmly closed. When the cartridge entrance
port on the top of the DiagCORE® Analyzer automatically opens, insert the cartridge with the
barcode facing to the left and the reaction chambers facing down.
There is no need to push the cartridge into the DiagCORE® Analyzer. Position it correctly into the
cartridge entrance port, and the DiagCORE® Analyzer will automatically move the cartridge
into the Analytical Module.
DiagCORE Respiratory Panel 2 Instructions for Use 23
8. Upon detecting the cartridge, the DiagCORE
®
Analyzer 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 DiagCORE® Analyzer will not accept a 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 pushing the Cancel
button in the bottom right corner of the screen.
NOTE
Depending on the system configuration, the operator may be required to
re-type their user password to start the test run.
NOTE
The lid of the cartridge entrance port will close automatically after 30
seconds if no cartridge is positioned in the port. If this occurs, repeat the
procedure starting with step 5.
9. While the test is running, the time remaining in the run is displayed on the screen.
24 DiagCORE Respiratory Panel 2 Instructions for Use
10. After the test run is completed, the Eject screen will appear and the Module Status Bar will show
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.
NOTE
If the test fails, refer to the DiagCORE Analyzer User Manual Troubleshooting
section for possible reasons and instructions on how to proceed.
11. Press Eject to remove the cartridge and dispose of it as biohazardous waste in accordance
with all national, state, and local health and safety regulations and laws.
The 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 be automatically moved back
into the Analyzer, and the lid of the cartridge entrance port will close. If this occurs, press Eject
to open the lid of the cartridge entrance port again and then remove the cartridge.
NOTE
Used cartridges must be discarded.
It is not possible to re-use cartridges for tests for which the execution was
started but then subsequently canceled by the operator, or for which an
error was detected.
12. After the cartridge has been ejected, the Results Summary screen will appear. Please refer to
section Interpretation of Results for further details. To run another test, press Run Test.
NOTE
For further information on the use of the instrument, refer to the DiagCORE
Analyzer User Manual.
DiagCORE Respiratory Panel 2 Instructions for Use 25
6 Procedure for transport medium liquid
samples
6.1 Sample collection, transport, and storage
1. Collect nasopharyngeal swab samples according to the swab manufacturer’s recommended
procedures, and place the swab into Universal Transport Medium.
6.2 Loading a sample into the DiagCORE
®
Respiratory Panel 2 Cartridge
1. Open the package of a DiagCORE
®
Respiratory Panel 2 Cartridge using the tear notches on
the sides of the packaging.
After the package is open, sample should be introduced inside the DiagCORE® Respiratory
Panel 2 cartridge and loaded in the DiagCORE® Analyser within 120 minutes.
2. Take out the cartridge and position it so that the QR code on the label faces you.
26 DiagCORE Respiratory Panel 2 Instructions for Use
3. Manually write the sample information or place a sample information label on the top of the
DiagCORE® cartridge. Make sure that the label is properly positioned and does not block the
lid opening.
4. Open the sample lid of the main port on the front of the cartridge.
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 (300 µL).
DiagCORE Respiratory Panel 2 Instructions for Use 27
IMPORTANT
Take care to not draw air into the pipette. If COPAN UTM 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.
6. Carefully transfer 300 µL of sample volume into the main port of the DiagCORE
®
Respiratory
Panel 2 cartridge, using a supplied single-use transfer pipette.
28 DiagCORE Respiratory Panel 2 Instructions for Use
7. Firmly close the lid of the main port until it clicks.
8. Confirm visually that the sample has been loaded by checking the sample inspection window
in the cartridge (see the image below).
After sample is introduced inside the DiagCORE® Respiratory Panel 2 cartridge, cartridge must
be loaded in the DiagCORE® Analyser within 90 minutes.
6.3 Starting the DiagCORE
®
Analyzer
1. Press the On/Off button on the front of the Analyzer to start the unit.
DiagCORE Respiratory Panel 2 Instructions for Use 29
NOTE
The power switch at the back of the Analytical Module must be set in the “I”
position. The DiagCORE® Analyzer status indicators turn blue in this case.
2. Wait until the Main screen appears and the DiagCORE
®
Analyzer status indicators turn green
and stop blinking.
3. Log in to the DiagCORE
®
Analyzer 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.
4. In case the Assay Definition File software has not been installed in the Analyzer follow the
instructions prior to running the test (See Section 9 of this document for additional information).
6.4 Running a test
1. Press the Run Test button at the top right corner of the touchscreen of the DiagCORE
®
Analyzer.
2. When prompted, scan the sample ID barcode on the UTM tube containing the sample, or scan
the specimen information barcode placed on the top of the DiagCORE® cartridge, using the
front barcode reader that is integrated into the DiagCORE® Analyzer.
NOTE
It is also possible to enter the sample ID using the virtual keyboard of the
touchscreen by pressing on the Sample ID field.
30 DiagCORE Respiratory Panel 2 Instructions for Use
NOTE
Depending on the chosen system configuration, entering the patient ID may
also be required at this point.
NOTE
Instructions from the DiagCORE® Analyzer appear in the Instructions Bar at
the bottom of the touchscreen.
3. When prompted, scan the barcode of the DiagCORE
®
Respiratory Panel 2 Cartridge to be used.
The DiagCORE® Analyzer automatically recognizes the assay to be run, based on the cartridge
barcode.
NOTE
The DiagCORE® Analyzer will not accept cartridges with elapsed expiration
date, previously used cartridges or cartridges for assays that are not
installed on the unit. An error message will be shown in these cases and the
cartridge will be rejected. Refer to the DiagCORE® Analyzer User Manual for
further details.
4. Select the appropriate sample type from the list.
5. The Confirm screen will appear. Review the data entered and make any necessary changes
by pressing the relevant fields on the touchscreen and editing the information.
6. Press Confirm when all the displayed data are correct. If needed, press on the appropriate field
to edit its content, or press Cancel to cancel the test.
DiagCORE Respiratory Panel 2 Instructions for Use 31
7. Ensure that both sample lids of the cartridge are firmly closed. When the cartridge entrance
port on the top of the DiagCORE® Analyzer automatically opens, insert the cartridge with the
barcode facing to the left and the reaction chambers facing down.
There is no need to push the cartridge into the DiagCORE® Analyzer. Position it correctly into the
cartridge entrance port, and the DiagCORE® Analyzer will automatically move the cartridge
into the Analytical Module.
8. Upon detecting the cartridge, the DiagCORE
®
Analyzer 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 DiagCORE® Analyzer will not accept a 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 pushing the Cancel
button in the bottom right corner of the screen.
NOTE
Depending on the system configuration, the operator may be required
to re-type their user password to start the test run.
32 DiagCORE Respiratory Panel 2 Instructions for Use
NOTE
The lid of the cartridge entrance port will close automatically after 30
seconds if no cartridge is positioned in the port. If this occurs, repeat the
procedure starting with step 5.
9. While the test is running, the time remaining in the run is displayed on the screen.
10. After the test run is completed, the Eject screen will appear and the Module Status Bar will show
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.
NOTE
If the test fails, refer to the DiagCORE Analyzer User Manual Troubleshooting
section for possible reasons and instructions on how to proceed.
11. Press Eject to remove the cartridge and dispose of it as biohazardous waste in accordance
with all national, state, and local health and safety regulations and laws.
The 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 be automatically moved back
into the Analyzer, and the lid of the cartridge entrance port will close. If this occurs, press Eject
to open the lid of the cartridge entrance port again and then remove the cartridge.
NOTE
Used cartridges must be discarded.
It is not possible to re-use cartridges for tests for which the execution was
started but then subsequently canceled by the operator, or for which an
error was detected.
12. After the cartridge has been ejected, the Results Summary screen will appear. Please refer to
section Interpretation of Results for further details. To run another test, press Run Test.
DiagCORE Respiratory Panel 2 Instructions for Use 33
NOTE
For further information on the use of the instrument, refer to the DiagCORE
Analyzer User Manual.
34 DiagCORE Respiratory Panel 2 Instructions for Use
7 Internal control
The DiagCORE® Respiratory Panel 2 Cartridge includes a full process Internal Control which is titered
MS2 bacteriophage. The MS2 bacteriophage is a single-stranded RNA virus, and it 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
DiagCORE® Respiratory Panel 2 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.
DiagCORE Respiratory Panel 2 Instructions for Use 35
8 Interpretation of results
8.1 Viewing results
The DiagCORE® Analyzer automatically interprets and saves test results. After ejecting the
cartridge, the Results Summary screen is automatically displayed.
Example screen showing Test Data on the left panel and Test Summary in the main panel.
The main part of the screen provides the following two 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 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.
Note that the pathogens detected and identified in the sample are shown in both lists.
In case 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
36 DiagCORE Respiratory Panel 2 Instructions for Use
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).
A report with the assay data can be exported to an external USB storage device. Insert the USB
storage device into one of the USB ports of the DiagCORE® Analyzer and press Save Report in the
bottom bar of the screen. This report can be exported later at any time by selecting the test from
the View Result List.
The report can also be sent to the printer by pressing Print Report in the bottom bar of the screen.
8.1.1 Viewing amplification curves
To view test amplification curves of pathogens detected, press the Amplification Curves tab.
Amplification Curves screen (Pathogens).
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 DiagCORE® Analyzer the Amplification
Curves screen is only available for operators who have 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 this pathogen.
Unselected pathogens will be shown in gray.
The corresponding CT and endpoint fluorescence values are shown below each pathogen name.
Press the Controls tab on the left side to view the controls and select which controls are shown in
the amplification plot. Press the circle next to the control name to select or deselect it.
DiagCORE Respiratory Panel 2 Instructions for Use 37
Amplification Curves screen (Pathogens and Controls).
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 gray pickers on each axis. Press and
hold a picker and then move it to the desired location on the axis. Move a picker to the axis
origin to return to the default values.
8.1.2 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:
User ID
Cartridge serial number
Cartridge expiration date
Analytical Module serial number
Test status (completed, failed, canceled by operator)
Error code if applicable
Test start date and time
Test execution time
Assay name
Test ID
Test Result: Positive (in case at least one respiratory pathogen is detected/identified), negative
(in case no respiratory pathogen is detected) or failed (an error occurred or the test was
canceled by the user)
38 DiagCORE Respiratory Panel 2 Instructions for Use
List of analytes tested in the assay, with C
T
and endpoint fluorescence in case of a positive signal
Internal Control, with C
T
and endpoint fluorescence
Example screen showing Test Data on the left panel and Test Details in the main panel.
8.1.3 Browsing results from previous tests
To view results from previous tests that are stored in the results repository, press View Results from
the Main Menu bar.
Example of View Results List.
DiagCORE Respiratory Panel 2 Instructions for Use 39
The following information is available for every executed test:
Sample ID
Assay Name
Operator ID
Analytical Module on which the test was executed
Date and time when the test was finished
Outcome of the test (positive (pos), negative (neg), failed (fail) or successful (suc))
NOTE
When User Access Control is enabled on the DiagCORE® Analyzer, the data
for which the user has no access rights will be hidden with asterisks.
Select one or more test results by pressing the 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 button in the top row.
Example of selecting test results in the View Results list.
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:
Name
Button
Description
Positive
pos
At least one pathogen is positive.
Negative
neg
No analytes were detected.
40 DiagCORE Respiratory Panel 2 Instructions for Use
Name
Button
Description
Failed
fail
The test failed, either because an error occurred or the test was
cancelled by the user.
Successful
suc
The test is either positive or negative, but the user does not have the
access rights to view the test results.
Please make sure a printer is connected to DiagCORE system and 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 the Search button 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.
The symbol to the left of a column headline indicates that the column’s filter is active.
A filter can be removed by pressing the Remove Filter button in the submenu bar.
8.1.4 Exporting results to a USB drive
From any tab of the test 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 instrument.
8.1.5 Printing results
Please make sure a printer is connected to DiagCORE system and proper driver is installed.
Select Print Report to send a copy of the test results to the printer.
8.2 Result interpretation
A result for a respiratory organism is interpreted as “Positive” when the corresponding PCR assay is
positive, except for Influenza A. The Influenza A assay in the DiagCORE® Respiratory Panel 2 is
designed to detect Influenza A as well as Influenza A subtype H1N1/2009, Influenza A subtype H1or
Influenza A subtype H3, respectively. In particular, this means:
In case seasonal Influenza A H1 strain is detected by the DiagCORE
®
Respiratory Panel 2
assay, two signals will be generated, one for Influenza A and a second one for H1 strain
and shown on the DiagCORE® Analyzer screen.
DiagCORE Respiratory Panel 2 Instructions for Use 41
In case seasonal Influenza A H3 strain is detected by the DiagCORE
®
Respiratory Panel 2
assay, two signals will be generated, one for Influenza A and a second one for H3 strain
and shown on the DiagCORE® Analyzer screen.
In case a pandemic Influenza A/H1N1/2009 strain is detected, two signals will be
generated, one for Influenza A and a second one for H1N1/2009 and shown on the
DiagCORE® Analyzer screen.
For every other pathogen that can be detected with the DiagCORE® Respiratory Panel 2, only one
signal will be generated if the pathogen is present in the sample.
8.3 Internal Control interpretation
Controls result
Explanation
Action
Passed
The internal control amplified
successfully.
The run was completed with success,
and all results are validated and can be
reported. Detected pathogens are
reported as positive, and not detected
pathogens are reported as negative.
Failed
The internal control failed.
Positively detected pathogen(s) are
reported, but all negative results (tested
but not detected pathogen(s)) are
invalid.
Repeat the testing using a new
DiagCORE® Respiratory Panel 2
Cartridge.
42 DiagCORE Respiratory Panel 2 Instructions for Use
9 Installing the Assay Definition File
The Assay Definition File of the DiagCORE® Respiratory Panel 2 has to be installed in the DiagCORE®
Analyzer previously to execute tests with the DiagCORE® Respiratory Panel 2 cartridges.
The same procedure has to be applied when a new version of the DiagCORE® Respiratory Panel 2
assay is released.
NOTE
Assay Definition Files are available on the STAT-Dx web site and also through the
STAT-Dx sellers/distributors.
The Assay Definition File (extension.asy) must be saved into a USB Drive previously
to the installation on the DiagCORE® Analyzer. This USB Drive must be formatted
with a FAT32 file system.
To import new assays from the USB to the DiagCORE® Analyzer proceed with the following steps:
1. Insert the USB stick with the Assay Definition File on one of the USB ports of the DiagCORE
®
Analyzer.
2. Press the Options button and then the Assay Management button. The Assay
Management screen appears on the content area of the display (See screenshot below).
DiagCORE Respiratory Panel 2 Instructions for Use 43
3. Press the Import icon in the bottom left of the screen.
4. Select the file from the USB drive corresponding to the assay to be imported.
5. A dialog will appear to confirm upload of the file.
6. A dialog may appear to override current version by a new one. Press yes to override.
7. The assay becomes active by pressing the ‘Assay Active’ button (See screenshot below).
44 DiagCORE Respiratory Panel 2 Instructions for Use
10 Limitations of the procedure
Results from the DiagCORE
®
Respiratory Panel 2 are not intended to be used as the sole basis
for the diagnosis, treatment, or other patient management decisions.
Positive results do not rule out co-infection with organisms not included in the DiagCORE
®
Respiratory Panel 2. 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.
A negative result with the DiagCORE
®
Respiratory Panel 2 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 DiagCORE
®
Respiratory Panel 2 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 transport medium, from individuals with
acute respiratory symptoms.
The DiagCORE
®
Respiratory Panel 2 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 DiagCORE
®
Respiratory Panel 2 must be interpreted by a trained healthcare
professional within the context of all relevant clinical, laboratory, and epidemiological findings.
The DiagCORE
®
Respiratory Panel 2 can be used only with the DiagCORE® Analyzer.
The DiagCORE
®
Respiratory Panel 2 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 material nucleic acids depends on proper sample collection, handling,
transportation, storage, and loading into the DiagCORE® Respiratory Panel 2 Cartridge.
Improper operations for any of the aforementioned can cause incorrect results, including false-
positive or false-negative results.
DiagCORE Respiratory Panel 2 Instructions for Use 45
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. Please note that a higher prevalence favors
the positive predictive value of a test results, while a lower prevalence favors the negative
predictive value of a test result.
46 DiagCORE Respiratory Panel 2 Instructions for Use
11 Performance characteristics.
Clinical Performance
The performance characteristics of the DiagCORE Respiratory Panel 2 assay were assessed in a
multicenter clinical trial. Both the performance of the universal transport medium of
nasopharyngeal swab specimen (´UTM´), and of a dry nasopharyngeal swab specimen
(FLOQSwabs, Copan ref 503CS01) (´SWAB´) were assessed. In the latter case, a swab is directly
entered in the DiagCORE cartridge after collection avoiding the transfer into a liquid medium. This
testing approach may greatly support safe and error-free sample managment, especially at the
point of care settting.
The study was designed as observational, prospective-retrospective, using left-over samples
obtained from subjects with signs and symptoms of an acute respiratory infection. Participating
sites were asked to test fresh and/or frozen clinical samples, according to a protocol and
site/specific instructions.
Three (3) hospital laboratories, located in Copenhagen (Denmark), Bonn, (Germany) and Paris,
(France) participated in the study. Samples tested by the DiagCORE Respiratory Panel 2 were
compared with the results of the standard of care (SOC) method(s) at the sites, as wells as with a
range of validated and commercially available molecular methods. This approach provided
results for pathogens not detected by SOC and/or allowed for final discrepancy resolution of
discordant results. As such the DiagCORE Respiratory Panel 2 assay results were compared against
Filmarray Respiratory Panel 1.7 & 2 and the Allplex Respiratory Panel assay.
A total of 578 clinical UTM patient samples were enrolled into the study. One (1) sample was
excluded from the analysis due to the sample being misplaced between DiagCORE and
comparator testing. Seven (7) of 577 samples failed initial testing, resulting in a first testing success
rate of 98.8%. The failure rate includes the failure rate of the internal control, which was 0.17%
(1/577). Two (2) samples could not be retested due to insufficient remaining specimen volume. The
sample that showed an initial internal control failure was successful upon retesting.
Fifteen (15) pathogen results could not be resolved because there was no SOC result (10 results) or
no resolution method result available (5 results). This resulted in the loss of 2 samples, the remaining
unresolvable results were in samples with multiple pathogens detected (coinfection samples).
Clinical sensitivity or Positive Percent Agreement (PPA) was calculated as 100% x (TP / TP + FN). True
positive (TP) indicates that both the DiagCORE Respiratory Panel 2 and comparator(s) methods
had a positive result for the organism, and false negative (FN) indicates that the DiagCORE
Respiratory Panel 2 result was negative while the comparator resolution methods results were
positive. Specificity or Negative Percent Agreement (NPA) was calculated as 100% x (TN / TN + FP).
True negative (TN) indicates that both the DiagCORE Respiratory Panel 2 and the comparator
method had negative results, and a false positive (FP) indicates that the DiagCORE Respiratory
Panel 2 result was positive but the comparator methods results were negative. For the calculation
of the clincial specificity of the individual pathogens the total available results were used, with the
DiagCORE Respiratory Panel 2 Instructions for Use 47
concerning true and false positive organism results subtracted. The exact binomial two-sided 95%
confidence interval was calculated for each point estimate.
A total of 698 results were available for analysis*. Overall Clinical Sensitivity or PPA could be
calculated from 475 results. The overall Clinical Specificity or NPA was calculated from 190 full
negative samples. In total 462 true positive and 204 true negative DiagCORE Respiratory Panel 2
results were found, as well as 13 false negative and 17 false positive results.
Table 1. DiagCORE Respiratory Panel 2 Clinical Sensitivity and Specificity or Positive Percent
Agreement and Clinical specificity or Negative Percent Agreement, with 95% Confidence
Intervals
DiagCORE Respiratory Panel 2
TP/
Sensitivity
/PPA
95% CI
TN/
Specificity
/NPA
95% CI
TP+FN
TN+FP
Overall
462/475
97.30%
95.4%-
98.4%
187/190
98.40%
95.5%-
99.5%
Viruses
Adenovirus
35/38
97.20%
85.8%-
99.5%
659/662
99.50%
98.7%-
99.,8%
Bocavirus
4/4
100.00%
51.0%-
100.0%
693/694
99.90%
99.2%-
100.0%
Coronavirus 229E
4/5
80.00%
37.6%-
96.4%
693/693
100.00%
99.4%-
100.0%
Coronavirus HKU1
8/8
100.00%
67.6%-
100.0%
690/690
100.00%
99.4%-
100.0%
Coronavirus OC43
10/10
100.00%
72.2%-
100.0%
688/688
100.00%
99.4%-
100.0%
Coronavirus NL63
22/24
91.70%
74.2%-
97.7%
674/674
100.00%
99.4%-
100.0%
Human
Rhinovirus/Enterovirus
56/59
94.9%
86.1%-
98.3%
629/639
98.40%
97.1%-
99.1%
Human
Metapneumovirus
22/22
100.00%
85.1%-
100.0%
676/676
100.00%
99.4%-
100.0%
Influenza A H3N2
36/36
100.00%
90.4%-
100.0%
662/662
100.00%
99.4%-
100.0%
Influenza A H1N1
29/29
100.00%
88.3%-
100.0%
669/669
100.00%
99.4%-
100.0%
Influenza A H1-2009
strain (pandemic)
11/12
91.70%
64.5%-
98.5%
688/688
100.00%
99.4%-
100.0%
Influenza B
55/56
98.20%
90.6%-
99.7%
642/642
100.00%
99.4%-
100.0%
Parainfluenza Virus 1
(PIV 1)
19/19
100.00%
83.2%-
100.0%
696/696
100.00%
99.5%-
100.0%
Parainfluenza Virus 2
(PIV 2)
3/3
100.00%
43.8%-
100.0%
695/695
100.00%
99.5%-
100.0%
Parainfluenza Virus 3
(PIV 3)
9/9
100.00%
70.1%-
100.0%
689/689
100.00%
99.4%100,0%
Parainfluenza Virus 4
(PIV 4)
5/6
83.30%
43.6%-
97.0%
691/692
99.90%
99.2%-
100.0%
Respiratory Syncytial
Virus
100/103
97.10%
91.8%-
99.0%
595/595
100.00%
99,4%-
100.0%
48 DiagCORE Respiratory Panel 2 Instructions for Use
DiagCORE Respiratory Panel 2
TP/
Sensitivity
/PPA
95% CI
TN/
Specificity
/NPA
95% CI
TP+FN
TN+FP
Bacteria
Bordetella pertussis
29/29
100.00%
88.3%-
100.0%
693/693
100.00%
99.4%-
100.0%
Mycoplasma
pneumoniae
21/21
100.00%
84.5%-
100.0%
676/677
99.80%
99.2%-
100.0%
*Note: There were 7 Chlamydophila pneumoniae pathogens found by the comparator methods
in the study samples. These were all correctly detected by the DiagCORE Respiartory Panel 2 but
are not subject to this CE mark and the sensitivity performance is therefore not reported. The 7
results were however incuded in the specificity calculation for the indiviual panel pathogens.
Note: there were no evaluable results available for Legionella pneumophila, both because this
pathogen was found in a low number in the study (2 detections), and because of the absence of
comparator method results.
Note: the sensitivity and specificity performance results for Parainfluenza Virus 1 (17 of 19 results)
and for Bordetella Pertussis (24 of 29 results) include results from a previous study (DiagCORE
Respiratory Panel assay study). This is a true reflection of the performance for these pathogens
because no design or other changes were made for these pathogens between these 2 assays.
Except for the sensitivity and specificity calculation of these respective organisms, these 44 results
are not part of the 698 results used to calculate the specificity performance for the remaining
(DiagCORE Respiratory Panel 2 assay pathogens.
The DiagCORE Respiratory Panel 2 assay detected multiple organisms in 101 samples, for a total of
228 organism results. This represents 26.3 % of the total positive specimens (101/ 385). Eighty-two
samples were double infections, 15 were triple infections, and the remaining coinfection samples
had 4 (3) or more pathogens (1 sample had 7 pathogens).
11.1 Dry swab specimen
To assess the ability and the clinical performance characteristics of the dry swab specimen when
entered directly into the DiagCORE Respiratory Panel 2 cartridge, a total of 448 clinical samples
were tested. This testing was conducted in 2 of the 3 sites that participated in the performance
evaluation of the UTM specimen. The objective was to demonstrate equivalency between
performance characteristics of the dry swab and the UTM specimens.
One clinical site had requested and obtained IRB approval to enroll patients for this part of the
study. Patients consenting to participate in the study provided 2 nasopharyngeal swabs, one from
each nostril. One swab was transferred into UTM and the other swab was directly entered into the
cartridge. Ninety-eight (98) swab samples were enrolled following this approach. To augment the
number of dry swab results and to ensure that all DiagCORE Respiratory Panel 2 pathogens were
represented in the dry swab testing, an additional 350 swabs were dipped in UTM. Because each
swab holds approximately 0.1 ml of liquid after dipping, two swabs were simultaneously dipped in
UTM, and entered into the cartridge. For all SWAB specimen, the simultaneously tested UTM
specimen served as the comparator method.
DiagCORE Respiratory Panel 2 Instructions for Use 49
A minimum of 5 dry swab results were available for each DiagCORE Respirary Panel 2 pathogen.
Parainfluenza Virus 4 and Legionella pneumophila formed an exception as only 3 and 2 results
respectively were avaialble.
Similar as above , the Clinical sensitivity or PPA was calculated as 100% x (TP / TP + FN). True positive
(TP) indicates that both the dry swab and the UTM specimen had a positive result for a specific
organism, and false negative (FN) indicates that the dry swab result was negative whi le the UTM
specimen result was positive. Specificity or NPA was calculated as 100% x (TN / TN + FP). True
negative (TN) indicates that both the dry swab and UTM specimen had negative results, and a
false positive (FP) indicates that the dry swab result was positive but the UTM specimen result was
negative. The exact binomial two-sided 95% confidence interval was calculated for each point
estimate
For all dry swabs, a total of 440 results were available for analysis. Overall Clinical Sensitivity or PPA
could be calculated from 244 results. The overall Clinical Specificity or NPA was calculated from
196 results . In total 241 true positive and 188 true negative dry swab results were found, as well as
3 false negative and 8 false positive dry swab results.
Because the same sample is tested, the dipped swab may be considered to be most relevant with
regard to assessing equivalency of the dry swab with the UTM specimen performance. Testing of
the dry swab involves taking 2 samples from the same patient and although paired, a bias may
occur due to this approach. Also, as the nasopharyngeal swab collection does present a level of
discomfort for the patient it is likely that the yields obtained between the 2 collections may differ.
For all dipped swabs, a total of 337 results were available for analysis. Overall Clinical Sensitivity or
PPA could be calculated from 178 results. The overall Clinical Specificity or NPA was calculated
from 159 results . In total 177 true positive and 156 true negative dipped swab results were found,
as well as 1 false negative and 3 false positive dipped swab results.
Table 3. DiagCORE Respiratory Panel 2 Sensitivity and Specificity characteristics with 95%
Confidence Intervals for dry swab specimen
DiagCORE Respiratory Panel 2
TP/
Sensitivity
95% CI
TN/
Specificity
95% CI
TP+FN
TN+FP
All Dry Swabs
241/244
98.80%
96.4%-99.6%
188/196
95.90%
92.2%-97.9%
Dipped Swabs
177/178
99.40%
96.9%-99.9%
156/159
98.10%
94.6%-99.4%
11.2 Conclusion
This extensive multicentre study seeked to assess the performance of the UTM specimen as well as
the equivalency of the dry swab with the UTM specimen performance in the DiagCORE Respiratory
Panel 2 assay.
The overall Clinical Sensitivity of the UTM specimen was found to be 97.3% (95% confidence interval
95.4%-98.4%). The overall Clinical Specificity in 190 full negative samples was 98.4% % (95%
confidence interval 95.5%-99.5%).
50 DiagCORE Respiratory Panel 2 Instructions for Use
The overall Clinical Sensitivity of the dry swab specimen was found to be 98. 8% (95% confidence
interval 96.4%-99.6%). The overall Clinical Specificity for the dry swab specimen was 98.1% % (95%
confidence interval 94.6%-99.4%).
The dry swab study results supported the ability to test swabs entered directly in the DiagCORE
Respiratory Panel 2 cartridges as dry swabs. The dry swab specimen was found to show excellent
concordance with the UTM specimen, as best demonstrated by the overall agreement between
the UTM specimen and dipped swabs, which was 98.5% (95% confidence interval 97%-99.5 %).
DiagCORE Respiratory Panel 2 Instructions for Use 51
12 Performance characteristics.
Analytical Performance
12.1 Sensitivity (Limit of Detection)
The Analytical Sensitivity or Limit of Detection (LoD) is defined as the lowest concentration at which
≥ 95% samples tested generate a positive call.
The LoD of the DiagCORE® Respiratory Panel 2 was determined per analyte using selected strains
representing individual pathogens possible to detect with the DiagCORE® Respiratory Panel 2.
Simulated NPS sample matrix (cultured human cells in Copan UTM) was spiked with one or more
pathogens and tested in 20 replicates.
Individual LoD values for each DiagCORE® Respiratory panel 2 target is shown in the table below.
Table 1. LoD values obtained for the different respiratory target strains tested in the DiagCORE®
Respiratory panel 2.
Pathogen
Strain
Source
Concentration
Detection
rate
Influenza A H1N1
A/New Jersey/8/76
ATCC VR-897
28.1 CEID50/mL
20/20
A/Brisbane/59/07
Zeptometrix
0810244CFHI
0.04 TCID50/mL
19/20
A/New
Caledonia/20/99
Zeptometrix
0810036CFHI
4,6 TCID50/mL
19/20
Influenza A H3N2
A/Virginia/ATCC6/2012
ATCC VR-1811
0.4 PFU/mL
19/20
A/Wisconsin/67/2005
Zeptometrix
0810252CFHI
2.5 TCID50/mL
20/20
A/Port Chalmers/1/73
ATCC VR-810
791.1 CEID50/mL
20/20
Influenza A, subtype
H1N1/2009
A/Virginia/ATCC1/2009
ATCC VR-1736
2.6 PFU/mL
20/20
A/SwineNY/03/2009
Zeptometrix
0810249CFHI
14.1 TCID50/mL
20/20
Influenza B
B/Virginia/ATCC5/2012
ATCC VR-1807
0.08 PFU/mL
20/20
B/FL/04/06
ATCC VR-1804
34.8 CEID50/mL
19/20
B/Taiwan/2/62
ATCC VR-295
28.1 CEID50/mL
20/20
Coronavirus 229E
-
ATCC VR-740
0.3 TCID50/mL
20/20
Coronavirus OC43
-
ATCC-1558
0.1 TCID50/mL
20/20
Coronavirus NL63
-
Zeptometrix
0810228CFHI
0.01 TCID50/mL
20/20
Coronavirus HKU1
-
Zeptometrix
NATRVP-IDI
1/3001
19/20
Parainfluenza Virus
1
C35
ATCC VR-94
23.4 TCID50/mL
20/20
Parainfluenza Virus
2
Greer
ATCC VR-92
5.0 TCID50/mL
19/20
Parainfluenza Virus
3
C 243
ATCC VR-93
15.8 TCID50/mL
20/20
Parainfluenza Virus
4a
M-25
ATCC VR-1378
2.8 TCID50/mL
20/20
Respiratory
Syncytial Virus A
A2
ATCC VR-1540
2.8 TCID50/mL
20/20
52 DiagCORE Respiratory Panel 2 Instructions for Use
Pathogen
Strain
Source
Concentration
Detection
rate
Respiratory
Syncytial Virus B
9320
ATCC VR-955
0.02 TCID50/mL
20/20
Human
Metapneumovirus
Peru6-2003 (type B2)
Zeptometrix
0810159CFHI
1.1 TCID50/mL
19/20
hMPV-16, IA10-2003
Zeptometrix
0810161CFHI
3.0 TCID50/mL
20/20
Adenovirus
GB (Adenovirus B3)
ATCC VR-3
50.0 TCID50/mL
20/20
RI-67 (Adenovirus E4)
ATCC VR-1572
15.8 TCID50/mL
20/20
Adenoid 75 (Adenoviurs
C5)
ATCC VR-5
5.0 TCID50/mL
20/20
Adenoid 71 (Adenovirus
C1)
ATCC VR-1
5.0 TCID50/mL
19/20
Adenovirus C2
ATCC VR-846
28.1 TCID50/mL
20/20
Adenovirus C6
ATCC VR-6
505.6 TCID50/mL
20/20
Bocavirus
Clinical sample
-
>1.0 copies/mL
20/20
Enterovirus
/US/IL/14-18952
(Enterovirus D68)
ATCC VR-1824
50.0 TCID50/mL
19/20
Echovirus 6 (D-1 (Cox))
ATCC VR-241
0.001 TCID50/mL
19/20
Rhinovirus
1059 (Rhinovirus B14)
ATCC VR-284
28.1 TCID50/mL
20/20
HGP (Rhinovirus A2)
ATCC VR-482
0.3 TCID50/mL
19/20
11757 (Rhinovirus A16)
ATCC VR-283
8.9 TCID50/mL
20/20
Type 1A
ATCC VR-1559
5.0 TCID50/mL
20/20
Mycoplasma
pneumoniae
M129-B7
ATCC 29342
0.1 CFU/mL
20/20
Legionella
pneumophila
CA1
ATCC 700711
>0.01 CFU/mL
20/20
Bordetella pertussis
I028
ATCC BAA2707
>0.001 CFU/mL
20/20
A639
Zeptometrix
NATRVP-IDI
1/100001
19/20
1
Relative dilution from stock concentration.
12.2 Assay Robustness
The verification of robust assay performance was assessed by analyzing the Internal Control
performance in clinical nasopharyngeal swab samples. 30 individual nasopharyngeal swab
samples negative for all pathogens possible to detect were analysed with the DiagCORE®
Respiratory Panel 2.
All samples tested showed a positive result and valid performance for the Internal Control of the
DiagCORE® Respiratory Panel 2.
12.3 Exclusivity (Analytical Specificity)
Exclusivity study was carried out by in silico analysis and in vitro testing to assess the Analytical
Specificity of the DiagCORE® Respiratory Panel 2 for respiratory or non-respiratory organisms which
are not covered by the panel. These organisms included specimens which are related to, but
distinct from, respiratory panel organisms, or that could be present in specimens collected from the
DiagCORE Respiratory Panel 2 Instructions for Use 53
intended test population. Selected organisms are clinically relevant (colonizing the upper
respiratory tract or causing respiratory symptoms), are common skin flora or laboratory
contaminants, or are microorganisms for which much of the population may have been infected.
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, preferably 105 TCID50/mL for viral targets and 106 CFU/mL for bacterial targets.
A certain level of cross-reactivity with Bordetella species was predicted by preliminary sequence
analysis and was observed when high concentrations of Bordetella holmesii were tested. No cross-
reactivity was observed with Bordetella bronchiseptica and Bordetella parapertussis at high
concentrations. Target gene used for Bordetella pertussis detection (insertion element IS481) is a
transposon also present in other Bordetella species. Table 1 below shows the list of pathogens
tested in this study.
Table 1. List of Analytical Specificity pathogens tested.
Type
Pathogen
Bacteria
Bordetella bronchiseptica
Bordetella holmesii
Bordetella parapertussis
Chlamydia trachomatis
Enterobacter aerogenes
Escherichia coli (0157)
Haemophilus influenzae
Klebsiella oxytoca
Klebsiella pneumoniae
Lactobacillus acidophilus
Moraxella catarrhalis
Mycoplasma genitalium
Mycoplasma hominis
Neisseria elongata
Neisseria gonorrhoeae
Neisseria meningitidis
Pseudomonas aeruginosa
Serratia marcescens
Staphylococcus aureus
Staphylococcus epidermidis
Stenotrophomonas maltophilia
Streptococcus agalactiae
Streptococcus pneumoniae
Streptococcus pyogenes
Streptococcus salivarus
Viruses
Cytomegalovirus
Epstein-Barr Virus
Herpes Simplex Virus 1
Herpes Simplex Virus 2
Measles Virus
Mumps
54 DiagCORE Respiratory Panel 2 Instructions for Use
Type
Pathogen
Fungi
Aspergillus fumigatus
Candida albicans
Cryptococcus neoformans
All pathogens tested showed a negative result, no cross-reactivity was observed for the organisms
tested in the DiagCORE® Respiratory Panel 2 (except for Bordetella holmesii as described above).
In silico analysis was performed for all primer/probe designs included in the DiagCORE® Respiratory
Panel 2 proving specific amplification and detection of targets without cross-reactivity.
12.4 Inclusivity (Analytical Reactivity)
An inclusivity study was performed to analyse the detection of a variety of strains that represent
the genetic diversity of each respiratory panel target organism (“inclusivity strains”). Inclusivity
strains for all analytes were included in the study, representative of the species/types for the
different organisms, e.g. a range of Influenza A strains isolated from different geogracphical areas
and in different calendar years were included. Next table below shows the list of respiratory
pathogens tested in this study.
Table 1. List of Analytical Reactivity pathogens tested.
Pathogen
Subtype/
Serotype
Strain
Source
Influenza A
H1N1
A/PR/8/34
ATCC VR-1469
A/New Jersey/8/76
ATCC VR-897
A/Brisbane/59/07
Zeptometrix 0810244CFHI
A/New Caledonia/20/99
Zeptometrix 0810036CFHI
H3N2
A/Virginia/ATCC6/2012
ATCC VR-1811
A/Wisconsin/67/2005
Zeptometrix 0810252CFHI
A/Port Chalmers/1/73
ATCC VR-810
A/Victoria/3/75
ATCC VR-822
A/Brisbane/10/07
Zeptometrix NATRVP-IDI
H1N1
(pan)
A/Virginia/ATCC2/2009
ATCC VR-1737
A/Virginia/ATCC3/2009
ATCC VR-1738
A/Virginia/ATCC1/2009
ATCC VR-1736
A/SwineNY/03/2009
Zeptometrix 0810249CFHI
H1N1/NY/02/09
Zeptometrix NATRVP-IDI
Influenza B
n/a
B/Virginia/ATCC5/2012
ATCC VR-1807
B/FL/04/06
ATCC VR-1804
B/Taiwan/2/62
ATCC VR-295
B/Panama/45/90
Zeptometrix NATFLUB-ERCM
B/Florida/02/06
Zeptometrix 810037CFHI
B/Maryland/1/59
ATCC VR-296
Coronavirus 229E
n/a
n/a
ATCC VR-740
n/a
Zeptometrix NATRVP-IDI
DiagCORE Respiratory Panel 2 Instructions for Use 55
Pathogen
Subtype/
Serotype
Strain
Source
Coronavirus OC43
n/a
n/a
ATCC-1558
n/a
Zeptometrix 0810024CFHI
n/a
Zeptometrix NATRVP-IDI
Coronavirus NL63
n/a
n/a
Zeptometrix 0810228CFHI
n/a
Zeptometrix NATRVP-IDI
Coronavirus HKU1
n/a
n/a
Zeptometrix NATRVP-IDI
Parainfluenza 1
n/a
C35
ATCC VR-94
n/a
Zeptometrix NATPARA1-ST
n/a
Zeptometrix NATRVP-IDI
Parainfluenza 2
n/a
Greer
ATCC VR-92
n/a
Zeptometrix 0810015CFHI
n/a
Zeptometrix NATRVP-IDI
Parainfluenza 3
n/a
C 243
ATCC VR-93
n/a
Zeptometrix NATPARA3-ST
n/a
Zeptometrix NATRVP-IDI
Parainfluenza 4
A
M-25
ATCC VR-1378
B
CH 19503
ATCC VR-1377
B
n/a
Zeptometrix NATRVP-IDI
RSV A
n/a
A2
ATCC VR-1540
Long
ATCC VR-26
n/a
Zeptometrix NATRVP-IDI
RSV B
n/a
9320
ATCC VR-955
18537
ATCC VR-1580
WV/14617/85
ATCC VR-1400
n/a
Zeptometrix NATRSVB-ST
Human
Metapneumovirus
B1
Peru2-2002
Zeptometrix 0810156CFHI
B1
IA18-2003
Zeptometrix 0810162CFH
B1
Peru3-2003
Zeptometrix 0810158CFHI
B2
Peru6-2003
Zeptometrix 0810159CFHI
B2
Peru1-2002
Zeptometrix 0810157CFHI
A1
hMPV-16, IA10-2003
Zeptometrix 0810161CFHI
A1
IA3-2002
Zeptometrix 0810160CFHI
A2
IA14-2003
Zeptometrix 0810163CFH
Adenovirus B
B21
AV-1645 [128]
ATCC VR-256
B7
Gomen
ATCC VR-7
B3
GB
ATCC VR-3
B3
n/a
Zeptometrix NATADV3-ST
Adenovirus C
C1
Adenoid 71
ATCC VR-1
C2
n/a
ATCC VR-846
C5
Adenoid 75
ATCC VR-5
C6
n/a
ATCC VR-6
Adenovirus E
E4
RI-67
ATCC VR-1572
Bocavirus
n/a
n/a
Zeptometrix 0601178NTS
n/a
Zeptometrix MB-004
56 DiagCORE Respiratory Panel 2 Instructions for Use
Pathogen
Subtype/
Serotype
Strain
Source
Enterovirus A
EV-A71
EV-A71
Zeptometrix 0810236CFHI
Enterovirus B
E-11
Gregory
ATCC VR-41
E-30
Bastianni
ATCC VR-1660
CV-A9
Griggs
ATCC VR-1311
CV-B1
Conn-5
ATCC VR-28
CV-B2
Ohio-1
ATCC VR-29
CV-B3
Nancy
ATCC VR-30
E-17
CHHE-29
ATCC VR-47
n/a
Echovirus 6 (D-1 (Cox))
ATCC VR-241
Enterovirus C
CV-A21
Kuykendall [V-024-001-012]
ATCC VR-850
Enterovirus D
D68
US/IL/14-18952
ATCC VR-1824
EV-D68
US/MO/14-18947
ATCC VR-1823
Rhinovirus A
A1
n/a
Zeptometrix NATRVP-IDI
1A
n/a
ATCC VR-1559
A2
HGP
ATCC VR-482
A16
11757
ATCC VR-283
HRV-1B
B632
ATCC VR-1645
HRV-A39
209
ATCC VR-340
Rhinovirus B
B14
1059
ATCC VR-284
M. pneumoniae
1
PI 1428
ATCC 29085
n/a
M129
Zeptometrix NATMPN(M129)ERCM
n/a
M129-B7
ATCC 29342
n/a
FH strain of Eaton Agent
[NCTC 10119]
ATCC 15531
L. pneumophila
n/a
CA1
ATCC 700711
Legionella pneumophila
subsp. Pneumophila/
169-MN-H
ATCC 43703
n/a
Zeptometrix 0601645NTS
subsp. Pneumophila /
Philadelphia-1
ATCC 33152
B. pertussis
n/a
I028
ATCC BAA-2707
A639
Zeptometrix NATRVP-IDI
18323 [NCTC 10739]
ATCC 9797
All pathogens tested showed positive results at the concentration tested.
DiagCORE Respiratory Panel 2 Instructions for Use 57
12.5 Co-Infections
A Co-infections study was performed to verify that multiple DiagCORE® Respiratory Panel 2 analytes
included in one nasopharyngeal swab sample can be detected by the DiagCORE® Respiratory
Panel 2.
High and low concentrations of different organisms were combined in one sample. Selection of
organisms was made based on relevance, prevalence and layout of the DiagCORE® Respiratory
Panel 2 cartridge (distribution of targets in different reaction chambers).
Analytes were spiked into simulated NPS sample matrix (cultured human cells in UTM) in high (50x
LoD concentration) and low concentrations (5x LoD concentration) and tested in different
combinations. Table below shows the combination of co-infections tested in this study.
Table 1. List of Co-infections combinations tested.
Pathogens
Strain
Concentration
Influenza A/H3N2
A/Virginia/ATCC6/2012
50xLoD
Adenovirus C5
Adenoid 75
5xLoD
Influenza A/H3N2
Adenovirus C5
A/Virginia/ATCC6/2012
Adenoid 75
5xLoD
50xLoD
Parainfluenza 3
C243
50xLoD
Influenza A/H1N1/2009
NY/03/09
5xLoD
Parainfluenza 3
Influenza A/H1N1/2009
C243
NY/03/09
5xLoD
50xLoD
Respiratory Syncytial Virus A
A2
50xLoD
Influenza B
B/FL/04/06
5xLoD
Respiratory Syncytial Virus A
Influenza B
A2
B/FL/04/06
5xLoD
50xLoD
Adenovirus C5
Adenoid 75
50xLoD
Rhinovirus B, Type HRV-B14
1059
5xLoD
Adenovirus C5
Rhinovirus B, Type HRV-B14
Adenoid 75
1059
5xLoD
50xLoD
Respiratory Syncytial Virus A
A2
50xLoD
Rhinovirus B, Type HRV-B14
1059
5xLoD
Respiratory Syncytial Virus A
Rhinovirus B, Type HRV-B14
A2
1059
5xLoD
50xLoD
Respiratory Syncytial Virus B
9320
50xLoD
Bocavirus
n/a
5xLoD
Respiratory Syncytial Virus B
Bocavirus
9320
n/a
5xLoD
50xLoD
Coronavirus OC43
n/a
50xLoD
Rhinovirus B, Type HRV-B14
1059
5xLoD
Coronavirus OC43
Rhinovirus B, Type HRV-B14
n/a
1059
5xLoD
50xLoD
Human Metapneumovirus B2
Peru6-2003
50xLoD
Parainfluenza 1
C-35
5xLoD
Human Metapneumovirus B2
Parainfluenza 1
Peru6-2003
C-35
5xLoD
50xLoD
Coronavirus 229E
n/a
50xLoD
Respiratory Syncytial Virus A
A2
5xLoD
Coronavirus 229E
Respiratory Syncytial Virus A
n/a
A2
5xLoD
50xLoD
Respiratory Syncytial Virus B
9320
50xLoD
Coronavirus NL63
n/a
5xLoD
Respiratory Syncytial Virus B
9320
5xLoD
Coronavirus NL63
n/a
50xLoD
58 DiagCORE Respiratory Panel 2 Instructions for Use
All co-infections tested gave a positive result for the two pathogens combined at low and high
concentrations. No effect in results are observed due to the presence of co-infections in a sample
tested with the DiagCORE® Respiratory Panel 2.
12.6 Interfering Substances
The influence of potential interfering substances on the performance of the DiagCORE® Respiratory
Panel 2 was evaluated in this study. The interfering substances include endogenous as well as
exogenous substances that are normally found in the nasopharynx or may be introduced into NPS
specimens during specimen collection, respectively.
A set of selected samples which cover all the respiratory pathogens from the panel were used for
the Interfering Substances testing. Interfering substances were spiked into the selected samples at
a level predicted to be above the concentration of the substance likely to be found in an
authentic nasopharyngeal swab specimen. The selected samples were tested with and without
addition of the potential inhibitory substance for direct sample-to-sample comparison. Additionally
pathogen-negative samples were spiked with the potential inhibitory substances.
None of the tested substances showed interference with the Internal Control or the pathogens
included in the combined sample.
Table 1. Concentrations of the interfering substances tested for the DiagCORE® Respiratory Panel
2.
Endogenous substances
Concentration tested
Human genomic DNA
50 ng/µL
Human whole Blood
10% v/v
Human Mucin
0.5% v/v
Competitive microorganisms
Concentration tested
Staphylococcus aureus
(ATCC CRM-6538)
1.70E+08 CFU/mL
Streptococcus pneumoniae
(ATCC 6303)
1.25E+07 CFU/mL
Haemophilus influenzae
(ATCC 49766)
6.20E+08 CFU/mL
Candida albicans
(ATCC CRM-10231)
1.00E+06 CFU/mL
Herpes Simplex Virus 1
(ATCC VR-1789)
1.60E+07 TCID50/mL
Human Cytomegalovirus
(ATCC NATCMV-0005)
2.0E+04 TCID50/mL
Exogenous substances
Concentration tested
Utabon®
Nasal spray (decongestant)
10% v/v
RHINOmer®
Nasal spray (salt water solutions)
10% v/v
Tobramycin
6 mg/mL
Mupirocin
2.5% w/v
DiagCORE Respiratory Panel 2 Instructions for Use 59
12.7 Carryover
A carryover study was performed to evaluate the potential occurrence of cross contamination
between consecutive runs when using the DiagCORE® Respiratory Panel 2 on the DiagCORE®
Analyzer.
Samples of simulated NPS matrix with alternating high-positive and negative samples were
conducted on one DiagCORE® Analyzer.
No carryover between samples was observed in the DiagCORE® Respiratory Panel 2.
12.8 Reproducibility
To prove reproducible performance of the DiagCORE® Respiratory Panel 2 on the DiagCORE®
Analyzer, a set of selected samples composed of low concentrated analytes (3xLoD and 1xLoD)
and negative samples were tested. Samples were tested in replicates using different lots of
DiagCORE® Respiratory Panel 2 cartridges and tests were executed on different DiagCORE®
Analyzers by different operators on different days.
Table 1: List of respiratory pathogens tested in Reproducibility
Pathogen
Strain
Influenza A H1
A/New Jersey/8/76
Influenza A H3
A/Virginia/ATCC6/2012
Influenza A H1N1 pdm
A/SwineNY/03/2009
Influenza B
B/FL/04/06
Cor 229E
n/a
Cor OC43
n/a
Cor NL63
n/a
Cor HKU1
n/a
Parainfluenza Virus 1
C35
Parainfluenza Virus 2
Greer
Parainfluenza Virus 3
C 243
Parainfluenza Virus 4a
M-25
Rhinovirus
A16
Enterovirus
/US/IL/14-18952 (enterovirus D68)
Adenovirus
RI-67 (adenovirus E4)
RSV B
9320
hMPV
Peru6-2003 (type B2)
Bocavirus
Clinical sample
Mycoplasma pneumoniae
M129-B7 (type 1)
Clamydophila pneumoniae
TW183
Legionella pneumophila
CA1
Bordetella pertussis
I028
60 DiagCORE Respiratory Panel 2 Instructions for Use
Table 2. Summary of Positive Agreement, Negative Agreement for Reproducibility Testing
Concentration
Pathogen
Expected
Result
Detection
Rate
% Agreement with
Expected Result
3xLoD
Influenza A H1 *
Positive
20/20
100%
Cor HKU1
Positive
20/20
100%
PIV2
Positive
20/20
100%
RSVB
Positive
20/20
100%
1xLoD
Influenza A H1 *
Positive
20/20
100%
Cor HKU1
Positive
19/20
95%
PIV2
Positive
19/20
95%
RSVB
Positive
20/20
100%
Negative
Influenza A H1 *
Negative
80/80
100%
Cor HKU1
Negative
80/80
100%
PIV2
Negative
80/80
100%
RSVB
Negative
80/80
100%
* Note: detection rate applies for both targets, Influenza A and H1.
Concentration
Pathogen
Expected
Result
Detection
Rate
% Agreement with
Expected Result
3xLoD
Influenza B
Positive
20/20
100%
Cor 229E
Positive
20/20
100%
PIV4a
Positive
20/20
100%
Enterovirus D68
Positive
20/20
100%
hMPV B2
Positive
20/20
100%
B. pertussis
Positive
20/20
100%
1xLoD
Influenza B
Positive
19/20
95%
Cor 229E
Positive
20/20
100%
PIV4a
Positive
20/20
100%
Enterovirus D68
Positive
19/20
95%
hMPV B2
Positive
19/20
95%
B. pertussis
Positive
20/20
100%
Negative
Influenza B
Negative
80/80
100%
Cor 229E
Negative
80/80
100%
PIV4a
Negative
80/80
100%
Enterovirus D68
Negative
80/80
100%
hMPV B2
Negative
80/80
100%
B. pertussis
Negative
80/80
100%
DiagCORE Respiratory Panel 2 Instructions for Use 61
Concentration
Pathogen
Expected Result
Detection
Rate
% Agreement with
Expected Result
3xLoD
Influenza H1N1 (pdm) *
Positive
20/20
100%
Cor OC43
Positive
20/20
100%
PIV3
Positive
20/20
100%
Rhinovirus A16
Positive
20/20
100%
M. pneumoniae
Positive
20/20
100%
1xLoD
Influenza H1N1 (pdm) *
Positive
20/20
100%
Cor OC43
Positive
20/20
100%
PIV3
Positive
20/20
100%
Rhinovirus A16
Positive
20/20
100%
M. pneumoniae
Positive
20/20
100%
Negative
Influenza H1N1 (pdm) *
Negative
80/80
100%
Cor OC43
Negative
80/80
100%
PIV3
Negative
80/80
100%
Rhinovirus A16
Negative
80/80
100%
M. pneumoniae
Negative
80/80
100%
* Note: detection rate applies for both targets, Influenza A and H1/pandemic.
Concentration
Pathogen
Expected
Result
Detection
Rate
% Agreement with
Expected Result
3xLoD
Influenza A H3 *
Positive
20/20
100%
Cor NL63
Positive
20/20
100%
PIV1
Positive
20/20
100%
Adenovirus E4
Positive
20/20
100%
L. pneumophila
Positive
20/20
100%
1xLoD
Influenza A H3 *
Positive
19/20
95%
Cor NL63
Positive
20/20
100%
PIV1
Positive
20/20
100%
Adenovirus E4
Positive
20/20
100%
L. pneumophila
Positive
20/20
100%
Negative
Influenza A H3 *
Negative
80/80
100%
Cor NL63
Negative
80/80
100%
PIV1
Negative
80/80
100%
Adenovirus E4
Negative
80/80
100%
L. pneumophila
Negative
80/80
100%
* Note: detection rate applies for both targets, Influenza A and H3.
Concentration
Pathogen
Expected
Result
Detection
Rate
% Agreement with
Expected Result
3xLoD
Bocavirus
Positive
20/20
100%
1xLoD
Bocavirus
Positive
20/20
100%
Negative
Bocavirus
Negative
80/80
100%
All samples tested generated the expected result (95-100% agreement) showing reproducible
performance of the DiagCORE® Respiratory Panel 2.
62 DiagCORE Respiratory Panel 2 Instructions for Use
Reproducibility testing demonstrated that the DiagCORE® Respiratory Panel 2 running in the
DiagCORE® Analyzer provides highly reproducible test results when the same samples are tested in
multiple runs, on multiple days, and with various operators using different DiagCORE® Analyzers and
multiple lots of cartridges.
12.9 Sample Stability
A sample stability study was executed to analyze storage conditions for clinical samples to be
tested with the DiagCORE® Respiratory Panel 2. Simulated NPS sample matrix (cultured human
cells in Copan UTM) was spiked with viral or bacterial culture material of low concentration (e.g. 3x
LoD). Samples were stored at the following conditions for testing:
15 ˚C to 25 ˚C for 4 hours
2 ˚C to 8 ˚C for 3 days
-15 ˚C to -25 ˚C for 30 days
-70 ˚C to -80 ˚C for 30 days
All pathogens were successfully detected at the different storage temperatures and durations
concluding that samples were stable at the indicated storage conditions and durations.
DiagCORE Respiratory Panel 2 Instructions for Use 63
13 References
1. Centers for Disease Control and Prevention (CDC). National Center for Immunization and
Respiratory Diseases (NCIRD). Division of Viral Diseases (DVD) web site.
2. World Health Organization. WHO Fact Sheet No221, November 2016. Influenza (seasonal).
http://www.who.int/mediacentre/factsheets/fs211/en/index.html. Accessed November 2016.
3. Flu.gov web site. About Flu. https://www.cdc.gov/flu/about/index.html.
4. Centers for Disease Control and Prevention (CDC). Diseases & Conditions: Human Parainfluenza
Viruses (HPIVs). https://www.cdc.gov/parainfluenza/index.html.
5. Centers for Disease Control and Prevention (CDC). Diseases & Conditions: Respiratory Syncytial
Virus Infection (RSV). https://www.cdc.gov/rsv/.
6. Centers for Disease Control and Prevention (CDC). Diseases & Conditions: Adenoviruses.
https://www.cdc.gov/adenovirus/index.html.
7. Centers for Disease Control and Prevention (CDC). Diseases & Conditions: Non-polio Enterovirus.
https://www.cdc.gov/non-polio-enterovirus/about/index.html.
8. Centers for Disease Control and Prevention (CDC). Diseases & Conditions: Mycoplasma
pneumoniae Infection. https://www.cdc.gov/pneumonia /atypical/mycoplasma /index.html.
9. Centers for Disease Control and Prevention (CDC). Diseases & Conditions: Pertussis (Whooping
Cough). https://www.cdc.gov/pertussis/
10. Protection of Laboratory Workers from Occupationally Acquired Infections; Approved
Guideline M29, Clinical Laboratory Standard Institute.
11. BLAST: Basic Local Alignment Search Tool. https://blast.ncbi.nlm.nih.gov/Blast.cgi
12. Schreckenberger PC and McAdam AJ, Point-Counterpoint: Large Multiplex PCR Panels Should
be First-Line Tests for Detection of Respiratory and Intestinal Pathogens. J Clin Microbiol, 2015
Oct; 53(10):3110-5.
64 DiagCORE Respiratory Panel 2 Instructions for Use
14 Appendices
14.1 Disposal information
Dispose in sealed containers at licensed waste disposal site. Dispose of contents/container
according with local/regional/national/international regulations.
14.2 Declaration of conformity
Name and address of the legal manufacturer:
STAT-Dx Life, S.L.
Baldiri Reixac 4-8
08028 Barcelona, Spain
An up-to-date Declaration of Conformity can be requested from STAT-Dx Life, S.L.
14.3 Glossary
Amplification curve: Graphical representation of the RT-real time PCR amplification data
Analytical Module (AM): The main DiagCORE® Analyzer hardware module, in charge of executing
tests on DiagCORE® cartridges. It is controlled by the Operational Module. Several Analytical
Modules can be connected to one Operational Module.
DiagCORE® Analyzer: The DiagCORE® Analyzer consists of an Operational Module and an
Analytical Module. The Operational Module includes elements that provide connectivity to the
Analytical Module and enable user interaction with the DiagCORE Analyzer. The Analytical Module
contains the hardware and software for sample testing and analysis.
DiagCORE® Respiratory Panel 2 cartridge: A self-contained disposable plastic device with all pre-
loaded reagents required for the complete execution of fully automated molecular assays for the
detection of respiratory pathogens.
IFU: Instructions For Use.
Main port: In the DiagCORE® respiratory panel 2 cartridge, inlet for transport medium liquid samples.
Nucleic acids: Biopolymers, or small biomolecules composed of nucleotides, which are monomers
made of three components; a 5-carbon sugar, a phosphate group and a nitrogenous base.
Operational Module (OM): The dedicated DiagCORE® Analyzer hardware that provides the user
interface for one to four Analytical Modules (AM).
PCR: Polymerase Chain Reaction
RT: Reverse Transcription
Swab port: In the DiagCORE® Respiratory Panel 2 cartridge, inlet for dry swabs.
User: A person who operates the DiagCORE® Analyzer/ DiagCORE® Respiratory Panel 2 cartridge
in the intended way.
DiagCORE Respiratory Panel 2 Instructions for Use 65
14.4 Trademarks and copyright
DiagCORE® is a registered trademark of STAT-Dx Life, S.L. Windows® is a registered trademark of
Microsoft Corporation.
All other trademarks are the property of their respective owners.
This DiagCORE® Respiratory Panel 2 cartridge is protected by copyright. Neither these Instructions
for Use, nor any part thereof, may be reproduced, modified, or distributed by any means now
known or later developed.
© 2017 STAT-Dx Life, S.L. — ALL RIGHTS RESERVED
14.5 Copyright, disclaimer, and warranty
INFORMATION IN THIS DiagCORE® Respiratory Panel 2 INSTRUCTIONS FOR USE IS PROVIDED IN
CONNECTION WITH STAT-Dx Life DiagCORE® Respiratory Panel 2 cartridge PRODUCT. NO LICENSE,
EXPRESS OR IMPLIED, BY ESTOPPEL OR OTHERWISE, TO ANY INTELLECTUAL PROPERTY RIGHTS IS
GRANTED BY THIS DOCUMENT. EXCEPT AS PROVIDED IN STAT-Dx Life TERMS AND CONDITIONS OF
SALE FOR THE DiagCORE® Respiratory Panel 2 cartridge, STAT-Dx Life ASSUMES NO LIABILITY
WHATSOEVER AND DISCLAIMS ANY EXPRESS OR IMPLIED WARRANTY RELATING TO THE USE OF THE
DiagCORE® Respiratory Panel 2 cartridge INCLUDING LIABILITY OR WARRANTIES RELATING TO
MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE, OR INFRINGEMENT OF ANY PATENT,
COPYRIGHT, OR OTHER INTELLECTUAL PROPERTY RIGHT ANYWHERE IN THE WORLD.
14.6 Patent statement
TO THE EXTENT PERMITTED BY LAW, NO RIGHTS FOR RESALE ARE CONFERRED WITH THE PURCHASE OF
THIS PRODUCT.
Purchase of the DiagCORE Analyzer and/or associated DiagCORE® Respiratory Panel 2 cartridges
does not grant any rights under patents owned or controlled, now or later, by STAT-Dx Life except
to the extent necessary to operate the DiagCORE Analyzer as specified in this DiagCORE Analyzer
User Manual or the DiagCORE® Respiratory Panel 2 cartridge as specified in this DiagCORE®
Respiratory Panel 2 cartridge Instructions for Use.
14.7 Software license agreement
Stat-Dx Life, S.L. is the owner, or licensee with right to sublicense, of all rights necessary to provide
to the Purchaser of the DiagCORE Analyzer and associated DiagCORE cartridges, a license to the
software embedded within the DiagCORE Analyzer or associated DiagCORE cartridges, software
supplied on physical media by STAT-Dx Life, or software downloaded to the DiagCORE Analyzer or
associated standard PC (see Glossary, section 14.3 above), collectively the “Software.” Physical
media and copies thereof in any form are, and remain, the property of STAT-Dx Life. The Software
is on loan to the Purchaser of the DiagCORE Analyzer and associated DiagCORE cartridges for the
term of this Software License Agreement. Purchaser must not cause or permit the decompilation,
disassembly, or reverse engineering of the Software; nor transfer the Software in whole or in part to
any third party without the prior written consent of STAT-Dx Life, which will not be unreasonably
withheld.
Stat-Dx Life grants to Purchaser of the DiagCORE Analyzer and associated DiagCORE cartridges a
non-exclusive, non-transferable license to use one copy of the Software in connection with one
66 DiagCORE Respiratory Panel 2 Instructions for Use
DiagCORE Analyzer, and to make one back-up copy. This Software License Agreement is effective
until terminated, and may be terminated by Stat-Dx Life if Purchaser of the DiagCORE Analyzer
and associated DiagCORE cartridges fails to comply with the terms and conditions of this Software
License Agreement, and is given written notice by Stat-Dx Life. Upon termination of this Software
License Agreement the Purchaser of the DiagCORE Analyzer and associated DiagCORE cartridges
must destroy all copies of the Software.
TO THE EXTENT NOT PROHIBITED BY LAW, IN NO EVENT SHALL STAT-Dx Life BE LIABLE FOR PERSONAL
INJURY OR ANY INCIDENTAL, SPECIAL, INDIRECT, OR CONSEQUENTIAL DAMAGES WHATSOEVER,
INCLUDING, WITHOUT LIMITATION, DAMAGES FOR LOSS OF PROFITS, LOSS OF DATA, BUSINESS
INTERRUPTION, OR ANY OTHER COMMERCIAL DAMAGES OR LOSSES, ARISING OUT OF OR RELATED
TO YOUR USE OF OR INABILITY TO USE THE SOFTWARE, HOWEVER CAUSED, REGARDLESS OF THE
THEORY OF LIABILITY (CONTRACT, TORT, OR OTHERWISE) AND EVEN IF LICENSOR HAS BEEN ADVISED
OF THE POSSIBILITY OF SUCH DAMAGES. SOME JURISDICTIONS DO NOT ALLOW THE LIMITATION OF
LIABILITY FOR PERSONAL INJURY, OR OF INCIDENTAL OR CONSEQUENTIAL DAMAGES, SO THIS
LIMITATION MAY NOT APPLY TO YOU. In no event shall STAT-Dx Life S.L.’s total liability to you for all
damages (other than as may be required by applicable law in cases involving personal injury)
exceed the purchase price of the DiagCORE Analyzer. The foregoing limitations will apply even if
the above stated remedy fails of its essential purpose.
To the extent that any terms and conditions in this DiagCORE® Respiratory Panel 2 Instructions for
Use differ from the terms and conditions in the Purchase or Sale Agreement for the DiagCORE®
Respiratory Panel 2, the terms and conditions in the Purchase or Sale Agreement will control.
DiagCORE® Respiratory Panel 2 Instructions for Use, Revision 1.0
© 2018 STAT-Dx Life S.L. All rights reserved.
STAT-Dx · Baldiri Reixac 4, Barcelona, Spain +34 93 448 51 24 · http.support.qiagen.com
DiagCORE®
Respiratory Panel 2
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