Agilent AriaDx Real-Time PCR User Manual

AriaDx Real-Time PCR System

Setup and User Guide

For In Vitro Diagnostic Use

Version F0, March 2021

Intended Use / Indications for Use

The AriaDx Real-Time PCR Dx instrument is a fully integrated quantitative PCR amplification, detection, and data analysis system for nucleic acids samples.

The instrument is to be used only by operators trained in laboratory techniques and procedures.

The customer is responsible for validation of assays and compliance with regulatory requirements that pertain to their procedures and uses of the instrument.

Notice

Each laboratory must validate their own assays for use on the AriaDx Real-Time PCR System. Assays must include controls designed to detect inconsistencies in instrument performance due to, for example, changes in optical performance or thermal block uniformity.

Symbol Table

European Conformity Caution

In Vitro Diagnostic Medical Device Catalog/code number

Manufacturer Consult Instructions for Use

Authorized representative in the European Community

Temperature limitation

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Content

Contents

1 Before You Begin

Overview of the AriaDx Real-Time PCR System 7 Materials provided with the AriaDx Real-Time PCR System 7 Hardware specifications 8 Recommended plasticware 8 Safety precautions 9 Indicator LED light 11 Agilent Technical Support 12

2 Installing and Setting Up the AriaDx Instrument

Installing the AriaDx Instrument 14

Step 1. Select a location for the instrument 14 Step 2. Unpack the shipping containers 14 Step 3. Install the optical modules 16 Step 4. Clean the thermal block 19 Step 5. Connect the instrument to a power supply 20 Step 6. Connect a keyboard or mouse to the instrument (optional) 20 Step 7. Connect the instrument to a network or directly to a PC 20

Setting Up the AriaDx Instrument 23

Step 1. Turn on the instrument and calibrate the background for the optical modules 23 Step 2. Set the instrument date and time 23 Step 3. Run a diagnostic check 24

Logging In and Creating User Accounts 25

Step 1. Log in to the instrument using an Administrator account 25 Step 2. Add user accounts 26 Step 3. Log in to your personal user account 26

Customizing Instrument Settings 27

3 Installing the Aria Software

Minimum requirements for running the Aria software 29

Installing the Standard Aria Software 30

Download the Aria software 30 Install the standard Aria software 30 Launch the Aria software 34

Installing the Aria ET (Electronic Tracking) Software 35

Download the Aria software and Microsoft SQL Server 2012 35

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Install Microsoft SQL Server 2012 35 Install the Aria ET software 39 Configure and start Microsoft Distributed Transaction Coordinator (MSDTC) service 44 Launch the Aria ET software 47

Installing Microsoft .NET Framework 4.0 49

Download Microsoft .NET Framework 4.0 49 Install Microsoft .NET Framework 4.0 49

4 Running Experiments

Preparing and Loading PCR Samples 53

Prepare samples 53 Load samples 53

Setting Up and Running Experiments 54

Use the touchscreen to set up and run an experiment 54 Use the PC software to set up and run an experiment 54

Managing Saved Experiments 56

Locate saved experiment files 56 Create a subfolder 56 Copy and paste experiments 57 Transfer experiments to a USB drive 57

5 Calibrating the AriaDx System

Calibrating the Instrument Motor 59

Calibrate the motor 59 Check the motor calibration date 59

Calibrating the Background 60

Calibrate the background 60 Check the background calibration status 60

Performing Other Calibration Procedures 61

Calibrate the touchscreen 61 Calibrate the analysis of high resolution melt data 61

6 Maintaining, Updating, and Disposing of the AriaDx System

Maintaining the Instrument 63

Service the instrument 63 Clean the instrument 63 Clean the optical modules 63

Updating the Instrument Software 65

Disposing of Instrument Parts 66

Cybersecurity Guidance 67

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7 Troubleshooting

Troubleshooting Instrument Error Messages 69

View error messages 69 Open the diagnostic report 69

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AriaDx Real-Time PCR System Setup and User Guide

1 Before You Begin

This chapter contains information for you to read and understand before you start setting up the instrument.

Overview of the AriaDx Real-Time PCR System

The AriaDx Real-Time PCR System is a fully integrated quantitative PCR amplification, detection, and data analysis system. The system design combines a thermal cycler, an optical system with an LED excitation source, and data analysis software. The instrument can hold up to six optics modules, and the scanning optics design delivers optimal separation between the dyes and between samples. The instrument provides a closed-tube PCR detection format that can be used with a variety of fluorescence detection chemistries including SYBR® Green and EvaGreen dyes as well as fluorogenic probe systems including TaqMan probes.

Materials provided with the AriaDx Real-Time PCR System

Table 1 Materials provided

Materials provided Quantity

AriaDx instrument 1

Certificate of Conformance 1

AriaDx installation poster 1

Power cord 1

Optical modules Up to 6 optical modules, as selected by the user

The AriaDx instrument, the Certificate of Conformance, and the AriaDx installation poster are all shipped together in the same shipping container. The power cord and optical modules are each shipped in their own packaging.

Table 2 Materials offered as options at time of purchase

Options Quantity

Electronic tracking (ET) software 1 seat

Notebook PC 1

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Hardware specifications

Table 3 Hardware specifications for the AriaDx Real-Time PCR System

Feature Specification

Excitation source Eight dye-specific LEDs per optical module

Detection Eight photodiodes per optical module

Dyes ROX, FAM, HEX, CY5, CY3, ATTO 425

Electrical power (input) 100–240 VAC, 50/60 Hz, 1100VA

Thermal system Peltier-based, 96-well block

Six slots, swappable cartridges, no reference channel needed

Thermal system temperature range

Cycling speeds 40 cycle protocol in 43 minutes

Storage environment 10–43°C (50–109°F); 10–90% non-condensing humidity

Operating environment 20–30°C (68–86°F); 20–80% non-condensing humidity;

Weight 50 lbs (23 kg)

Dimensions 19.7" W × 18.1" D × 16.5" H (50 cm × 46cm × 42cm)

On board quality checks Instrument QC checks:

Warranty One year warranty

Plastic consumables Low-profile 0.2-ml tubes, strip tubes, and 96-well plates

25.0–99.9°C Max Heating: >6°C/sec Max Cooling: >2.5°C/sec Accuracy: ±0.2°C or better at typical annealing, amplification, and denaturation temperatures

maximum altitude of 2000 m

All checks: 30 minutes; Selected checks: 10–30 minutes

Calibration: 1-minute background calibration

Data loss prevention: Data captured from all channels on each scan Ability to store up to 5 GB on the instrument

See “Recommended plasticware” on page 8 for part numbers

Recommended plasticware

Table Table 4 lists the part numbers of Agilent plates, tubes, and other plastic consumables for use in the AriaDx instrument.

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Table 4 QPCR plasticware for use in the AriaDx instrument

Agilent Part Number

401490 96-well plate, fully skirted, low profile

401491 96-well plate, rigid, fully skirted, low profile

401494 96-well plate, non-skirted, low profile

401493 8x strip tubes, without caps, low profile, 120 strips (10 packs of 12 strips)

401425 8x strip tube optical caps, 120 strips (10 packs of 12 strips)

401427 8x strip tube optical caps, 60 strips (5 packs of 12 strips)

401492 Adhesive seal for 96-well plates, 50-pack

Safety precautions

This product is designed for convenient and reliable operation, and to accepted standards of safety. Its use does not entail any hazard if operated in accordance with the instructions given in this document. However, incorrect operation may cause damage to the equipment or cause a hazard to health. It is important that the following safety precautions are read and understood before using the instrument.

Description

When sealing plates with adhesive seals, Agilent recommends using MicroAmp Optical Film Compression Pads (Life Technologies, part number 4312639).

All users must read and understand the Setup and User Guide and only operate the unit in accordance with the instructions given. Failure to follow instructions may cause the protection provided by the instrument to be impaired.

Electrical

Standard electrical safety precautions should be applied, including the following:

• Always put the instrument in a location where, if needed, the power supply can be immediately disconnected.

• Proper voltage (100–240 VAC) must be supplied before you turn on the instrument for the first time.

• The device must be connected to a grounded socket. Do not operate the instrument from a power outlet that has no ground connection.

• Do not touch any switches or outlets with wet hands.

• Turn off the instrument before you disconnect the power cord.

• Unplug the instrument before you clean any major liquid spills and before you service any of

the electrical or internal components.

• Do not connect the instrument to the same power strip as other high power-draw appliances (e.g., refrigerators and centrifuges).

• Do not service the electrical components unless you are qualified to do so.

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Fluids and Reagents

• Fill reaction vessels outside the instrument so that no fluids penetrate the instrument.

• Never cycle or incubate explosive, flammable and reactive substances in the instrument.

• You must observe the relevant safety regulations when handling pathogenic material,

radioactive substances or other substances hazardous to health.

• Do not submerge the instrument in any liquid.

Danger of Burns

• Do not touch the thermal block, inner side of heated lid and reaction vessels. These areas

quickly attain temperatures of greater than 50°C. Keep the heated lid closed until temperatures of 30°C or lower are reached.

• Do not use any materials (plates, sealings, foils, mats) which are not sufficiently temperature-stable (up to 120°C).

Operating Environment

• The ventilation slots of the device must remain free to vent at all times. Leave at least 10 cm of

space around the instrument.

• Keep the ambient temperature between 20°C and 30°C with humidity levels between 20% and 80% non-condensing.

• Do not operate the instrument in a hazardous or potentially explosive environment.

• Do not attempt to open the instrument door when the instrument is running an experiment.

Equipment Ratings

• Pollution degree 2

• Installation category II

• Altitude 2000 m

• Humidity 20 to 80%, non-condensing

• Electrical supply 100-240 VAC, 50/60 Hz, 1100VA

• Temperature 20°C to 30°C

• For Indoor Use Only

Electrostatic Discharge

The instrument is static sensitive. Electrostatic discharges greater than 8000 volts may interfere with the normal operation of the USB ports on the instrument. Handling precautions are required when working in high static environments. Wear a grounded wrist strap and take other antistatic precautions prior to making contact with the device in high static environments. ESD STM5.1-1998 Class 3B.

The AriaDx Real-Time PCR System complies to IEC 61326-2-6 standards that governs Electromagnetic Compliance for IVD medical equipment. This equipment has been designed and tested to CISPR 11 Class A. In a domestic environment it may cause radio interference, in which case, you may need to take measures to mitigate the interference. It is therefore advisable to choose a suitable electromagnetic environment for the AriaDx instrument prior to use.

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Safety Symbols

The electrical/safety symbols described below may be displayed on the instrument.

Indicator LED light

Power on

Power off

Caution

Caution, hot surfaces

On the front of the instrument (top right corner) is a status indicator LED. Table 5 summarizes the status codes for this LED light.

Table 5 Appearance of the status indicator LED

Appearance Instrument Status

Off The instrument is idle.

Blinking green The instrument is running.

Solid green The instrument is paused.

Blinking red The instrument has detected an error. Check the display for an error

message containing further details.

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Agilent Technical Support

Agilent Technical Support is available worldwide.

For US and Canada

Call (800)227-9770 (option 3,4,3)

Or send an e-mail to qPCR.support@agilent.com.

For all other regions

Agilent’s world-wide Sales and Support Center contact details for your location can be obtained at

www.agilent.com/en/contact-us/page.

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AriaDx Real-Time PCR System Setup and User Guide

2 Installing and Setting Up the AriaDx Instrument

Installing the AriaDx Instrument 14 Step 1. Select a location for the instrument 14 Step 2. Unpack the shipping containers 14 Step 3. Install the optical modules 16 Step 4. Clean the thermal block 19 Step 5. Connect the instrument to a power supply 20 Step 6. Connect a keyboard or mouse to the instrument (optional) 20 Step 7. Connect the instrument to a network or directly to a PC 20 Setting Up the AriaDx Instrument 23 Step 1. Turn on the instrument and calibrate the background for the optical modules 23 Step 2. Set the instrument date and time 23 Step 3. Run a diagnostic check 24 Logging In and Creating User Accounts 25 Step 1. Log in to the instrument using an Administrator account 25 Step 2. Add user accounts 26 Step 3. Log in to your personal user account 26 Customizing Instrument Settings 27

This chapter contains instructions for installing and setting up the AriaDx instrument.

Installing the AriaDx Instrument

Step 1. Select a location for the instrument

• Locate a solid, flat clean surface for the instrument. Make sure that:

• The instrument can stand completely stable.

• The rear air slots must not be covered.

• The instrument has at least 10 cm (approximately 4 inches) to the next wall or neighboring

instrument.

• The instrument is not located near anything that could be a source of vibrations.

• The temperature (normal ambient) is between 20°C and 30°C with humidity levels between

20% and 80% non-condensing.

• The atmosphere is not explosive.

Step 2. Unpack the shipping containers

NOTE

1 Open the small shipping container that holds the power cord. Remove the power cord and set

it aside for now.

2 Make sure that the large shipping container is in the upright position (Figure 1), then cut the

four plastic straps that hold the container together.

The AriaDx instrument is shipped in two separate containers. The small container holds the power cord. The large container holds the instrument and accessory tray.

Any optical modules that you ordered with the instrument are each packed and shipped separately in their own box.

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Figure 1 AriaDx instrument shipping container

3 Open the top flaps of the large container.

Inside the container is an accessory tray (see Figure 2), which contains the Certificate of Conformance and the installation poster.

Figure 2 Opened shipping container with accessory tray sitting on top

4 Remove the accessory tray. Unpack the installation poster from the accessory tray and use it

to guide you through the remainder of the unpacking process.

5 Remove the foam support that sits on top of the instrument in the shipping container.

6 Grip a handle on each side of the shipping container and lift up to remove the box sleeve that

surrounds the instrument. The instrument sits on the base of the shipping container, as shown in Figure 3.

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Figure 3 Instrument on shipping container base, with box sleeve removed

7 Remove the plastic wrapping from the instrument.

8 Lift the instrument off of the shipping container base and set it down on its selected location

(see “Step 1. Select a location for the instrument”).

Agilent recommends that two people lift the instrument together.

Step 3. Install the optical modules

1 Open the instrument door by lifting up on the handle on the top of the instrument. Lift the door

all the way up and back.

2 Remove the piece of foam and then remove the strip of cardboard from around the thermal

block assembly (see Figure 4). The optical module housing carrier is positioned to the left of the thermal block assembly.

Figure 4 Foam insert (left) and underlying cardboard strip (right).

3 Slide the optical module housing carrier to the right until it is centered in the opening of the

instrument door (as shown in Figure 5). Use the indentation on the top of the carrier to help slide it.

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Figure 5 Optical module housing carrier, centered in the instrument door opening

4 Open the lid on the optical module housing carrier.

a With your thumb and index finger, pinch together the two pieces of plastic in the indentation

on the top of the carrier (see Figure 6).

b Lift the lid all the way back to reveal the six slots for the optical modules (see Figure 7).

Figure 6 Opening of the optical module housing

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Figure 7 Slots for the optical modules

5 Open the boxes containing the optical modules. Remove the top piece of foam from each box

(Figure 8) then remove the plastic bag containing the optical module.

Figure 8 Optical module shipping box – top piece of foam removed

6 Install the optical modules into the slots.

a Open the plastic bag and remove the optical module.

b Peel off the plastic film from the edge of the optical module (see Figure 9). Once the film is

removed, do not to touch the exposed edge.

c Put the optical module into an available slot in the optical module housing. The correct

orientation for the optical module is label side up with the Agilent spark closer to the front of the instrument (see Figure 10).

NOTE

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If you are installing fewer than six optical modules, one or more of the slots in the optical module housing will be empty. In such cases, make sure that the empty slots are on the left-most side of the housing.

Figure 9 Removal of plastic film from optical module

Figure 10 Installed optical modules

7 Lower the lid on the optical module housing until it clicks shut.

When you turn on the instrument for the first time, it will prompt you to calibrate the background for the optical modules. Calibration is described in “Step 1. Turn on the instrument

and calibrate the background for the optical modules” on page 23.

Step 4. Clean the thermal block

With the instrument door still open, clean the outside and inside surfaces of the thermal block.

1 Lift the lid of the thermal block by pulling forward on the handle of the lid and then lifting the lid

up and away from the thermal block.

2 Using an aerosol can of compressed air, clean out the wells of the thermal block. Hold the can

8–10 cm away from the thermal block as you press the trigger.

3 Moisten a lint-free cleansing tissue with deionized H20, and gently wipe down the thermal

block and the underside of the lid. Then, close the lid of the thermal block and wipe down the top of the lid.

4 Close the instrument door.

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Step 5. Connect the instrument to a power supply

You must connect the instrument to a grounded AC outlet.

1 Plug the power cord into the power connector at the rear of the instrument.

See “Electrical” on page 9 for information on electrical safety precautions.

2 Connect the cable plug to the outlet.

Step 6. Connect a keyboard or mouse to the instrument (optional)

If desired, you can connect a keyboard or mouse to the instrument via the USB ports on the front and back of the instrument.

• Plug the USB cable of the device into a USB port on the instrument.

Multimedia keyboards are not supported.

Step 7. Connect the instrument to a network or directly to a PC

Connecting your instrument to a PC, either directly or though a network, allows you to remotely retrieve data from the instrument to your PC through the Aria PC software. If you do not connect your instrument to a PC or network, you must transfer post-run experiment data by copying it from the instrument to a USB drive (FAT format), and then from the USB drive to your PC.

To connect the instrument to a network

1 Plug one end of an ethernet cable into the ethernet port on the back of the instrument. Use a

standard Cat 6 straight/crossover ethernet cable.

2 Plug the other end of the cable into a network port.

To connect the instrument directly to a PC

1 Plug one end of an ethernet cable into the ethernet port on the back of the instrument. Use a

standard Cat 6 straight/crossover ethernet cable.

2 Plug the other end of the cable into the PC.

3 After you turn on the AriaDx instrument, set a static IP address, subnet mask, and default

gateway on the instrument using the instructions below. You will need to wait to complete these steps until after you complete “Step 1. Turn on the

instrument and calibrate the background for the optical modules” on page 23.

a On the Home screen of the AriaDx touchscreen, press Settings.

b Press Connection Settings. If you see an error message that no network connection is

detected, press OK to close the error message.

c On the Connection Settings screen, select Use Manual Configuration.

d In the IP Address, Subnet Mask, and Default Gateway fields, enter the values shown in

Figure 11. Press OK.

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Figure 11 Connection Settings

4 On the PC, set a static IP address, subnet mask, and default gateway using the instructions

below.

a Open the Control Panel to the Network and Sharing Center.

b Under View your active networks, click Local Area Connection (Figure 12). Note that the

appearance of the Network and Sharing Center screen may differ slightly from that shown in Figure 12 depending on your current network setup.

Figure 12 Network and Sharing Center – View your active networks

The Local Area Connection Status dialog box opens.

c In the Local Area Connection Status dialog box, click Properties.

The Local Area Connection Properties dialog box opens

d In the list of items under This connection uses the following items, double-click Internet

Protocol Version 4 (TCP/IPv4) (Figure 13).

Figure 13 Local Area Connection Properties dialog box – Internet Protocol Version 4

The Internet Protocol Version 4 (TCP/IPv4) Properties dialog box opens.

e On the General tab of the dialog box, select Use the following IP address and set the IP

address, subnet mask, and default gateway to the values shown in Figure 14. Click OK to close the dialog box.

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