Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a PCR-based diagnostic test and system. There is no mention of AI or ML in the intended use, device description, or performance studies. The analysis is based on detecting specific nucleic acids.

Therapeutic

No.
The device is a diagnostic test intended for the qualitative detection and identification of microbial nucleic acids, which aids in the diagnosis of respiratory infections. It does not provide any treatment or therapeutic benefit.

Diagnostic

Yes

The device detects and identifies multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs to aid in the diagnosis of respiratory infections. Both the "Intended Use / Indications for Use" and the "Device Description" sections clearly state its purpose in identifying pathogens and assisting in diagnosis.

SaMD (Software as a Medical Device)

No

The device description explicitly states that the QIAstat-Dx system is based on "single-test cartridges with pre-packaged reagents" and is processed by the "QIAstat-Dx® Analyzer 1.0". These are physical hardware components, not solely software.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The intended use explicitly states it's a "multiplexed nucleic acid test intended for use with QIAstat-Dx system for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs... obtained from individuals suspected of respiratory tract infections." This describes a test performed in vitro (outside the body) on a biological specimen (nasopharyngeal swab) to provide diagnostic information.
Device Description: The description details a system that processes a sample within a cartridge using pre-packaged reagents and PCR technology to detect analytes. This is characteristic of an in vitro diagnostic device.
Clinical Studies: The document describes clinical studies where the device's performance is evaluated by testing clinical specimens and comparing the results to a comparator method (an FDA-cleared multiplexed respiratory pathogen panel). This is a standard process for validating IVDs.
Key Metrics: The document provides performance metrics like Sensitivity/PPA and Specificity/NPA, which are key indicators used to evaluate the accuracy and reliability of diagnostic tests.
Predicate Device: The mention of a "Predicate Device(s)" (K123620; FilmArray® Respiratory Panel (RP)) is a strong indicator that this device is being submitted for regulatory clearance as an IVD, as predicate devices are used for comparison in the regulatory review process for new IVDs.

All these elements align with the definition and characteristics of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The QIAstat-Dx Respiratory Panel is a multiplexed nucleic acid test intended for use with QIAstat-Dx system for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) eluted in Universal Transport Media (UTM) obtained from individuals suspected of respiratory tract infections. The following organism types are identified using the QIAstat-Dx Respiratory Panel: Adenovirus, Coronavirus 229E, Coronavirus HKU1, Coronavirus NL63, Coronavirus OC43, Human Metapneumovirus A+B, Influenza A, Influenza A H1, Influenza A H1N1/pdm09, Influenza B, Parainfluenza Virus 1, Parainfluenza Virus 2, Parainfluenza Virus 4, Rhinovirus/Enterovirus, Respiratory Syncytial Virus A+B, Bordetella pertussis, Chlamydophila pneumoniae and Mycoplasma pneumoniae.

The detection and identification of specific viral and bacterial nucleic acids from individuals presenting with signs and symptoms of a respiratory infection aids in the diagnosis of respiratory infection with other clinical and epidemiological information. The results of this test should not be used as the sole basis for diagnosis, treatment or other management decisions. Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by the test or lower respiratory tract infection that is not detected by a nasopharyngeal swab specimen. Positive results do not rule out co-infection with other organisms: the agent(s) detected by the QIAstat-Dx Respiratory Panel may not be the definite cause of disease. Additional laboratory testing (e.g., bacterial and viral culture, immunofluorescence and radiography) may be necessary when evaluating a patient with possible respiratory tract infection.

Due to the small number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Bordetella pertussis and Parainfluenza Virus 1 were established primarily with retrospective clinical specimens. Performance characteristics for Chlamydophila pneumoniae, Parainfluenza Virus 2, Parainfluenza Virus 4, Influenza A subtype H1 and Coronavirus 229E were established primarily using contrived clinical specimens.

Due to the genetic similarity between Human Rhinovirus and Enterovirus, the QIAstat-Dx Respiratory Panel cannot reliably differentiate them. A positive QI Respiratory Panel Rhinovirus/Enterovirus result should be followed-up using an alternate method (e.g., cell culture or sequence analysis).

Performance characteristics for Influenza A were established when Influenza A H1N1-2009 and A H3 were the predominant Influenza A viruses in circulation. Performance of detecting Influenza A may vary if other Influenza A strains are circulating or a novel Influenza A virus emerges. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

Product codes (comma separated list FDA assigned to the subject device)

OCC, OEM, OOU, OEP, OTG, OQW, OOI, OZZ, OZY, OZX

Device Description

QIAstat-Dx is based on single-test cartridges with pre-packaged reagents including both wet and dry chemistry to handle the sample preparation and detection steps for the presence of a range of selected analytes by PCR technology. After insertion of the sample, the QIAstat-Dx assay cartridge is processed by the QIAstat-Dx Analyzer 1.0.

The OIAstat-Dx Respiratory Panel is intended to be used with liquid sample nasopharyngeal swabs (NPS).

Once the cartridge has been inserted into the instrument, the test starts automatically and runs for approximately 74 minutes. When the test is finished, the cartridge is removed by the user and discarded. The QIAstat-Dx Analyzer 1.0 automatically interprets test results and displays a summary on the analyzer display screen. The results can be printed using a connected printer if needed. The detected analytes are displayed in red. All other tested but not detected analytes are listed in green. The analyzer will report if an error occurs during processing, in which case the test will fail and no results will be provided (screen will show "FAIL").

Resuspension of IC and Prot K: Following insertion of the cartridge, the IC and Prot K are resuspended with the buffer located in Reservoir 1 (resuspension buffer from R1 is added to the interconnected IC cavity and Prot K cavity and transferred repeatedly between the Transfer Chamber and the cavities to ensure resuspension. The resuspended IC and Prot K are transferred to the sample cavity.

Cell Lysis: Primary lysis of the cells and analytes present in a NPS sample and IC occurs by a combination of chemical and mechanical processes using a rotor inside the lysis chamber in the presence of a buffer that acts as a chemical agent in aiding the mechanical process. The fast movement of the rotor results in sample agitation, which creates turbulence and shear forces that favor the lysis of the cell wall. After mechanical lysis is completed, the primary lysate is transferred to the purification chamber through a frit with 80 um pore size. The second lysis buffer (from Reservoir 2) is added to the primary lysate to complete chemical lysis.

Purification: Binding reagent (from Reservoir 4) is added to the lysate in the purification cavity, and the mix is passed through the silica membrane. In this process, the DNA/RNA molecules stick to the membrane, and the remaining components of the lysate are delivered to the waste chamber. Then the membrane is washed with a first washing buffer (from Reservoir 5) to wash away proteins. This is followed by a second washing step with a second washing buffer (from Reservoir 6) to remove any remaining substances other than the nucleic acids. A subsequent drying step eliminates volatile substances from the silica membrane. Prior to the elution step, the Transfer Chamber is rinsed with the rinsing buffer (from Reservoir 7) in order to remove any potential inhibitors from previous processing steps. At the end of the process, the nucleic acids are released from the membrane using an elution buffer (from Reservoir 8). The eluate is collected in the Transfer Chamber.

Rehydration of Master Mix: A defined volume (approximately 135uL) of the eluate is delivered to the dry chemistry reservoir (DCC) to rehydrate the Master Mix. Any remaining eluate is transferred to the waste chamber. The eluate/Master Mix solution is mixed by repeated transfer between the Transfer Chamber and the DCC.

Aliquotting and PCR: Defined aliquots (approximately 15 uL) of mixed eluate/Master Mix are sequentially transferred from the Transfer Chamber to each of seven Reaction Chambers containing the specified, air dried primers and probes. Within each Reaction Chamber, multiplex rtPCR testing is performed. Increase in fluorescence (indicative of detection of each target analyte) is detected directly within each Reaction Chamber. The rtPCR process is conducted by two submodules of the QIAstat-Dx Analyzer 1.0: the Thermal Cycler and the qPCR Sensor.

Components Description: QIAstat-Dx Respiratory Panel Cartridge: The QIAstat-Dx Respiratory Panel cartridge is a disposable plastic device that allows performing fully automated molecular assays. The main features of the QIAstat-Dx Respiratory Panel cartridge for the RP assay include the ability to test liquid samples as well as direct swabs and the capacity to store all necessary reagents within the cartridge needed for such testing. The cartridge is also designed to allow future expansion to incorporate additional sample types, such as swabs. All sample preparation and assay steps will be performed within the cartridge. All the reagents required for the complete execution of the test are pre-loaded and selfcontained in the QIAstat-Dx Respiratory Panel. The user does not need to manipulate any reagents. During the test, reagents are handled by pneumatically-operated microfluidics without any direct contact with the user or the analyzer actuators. This eliminates any possibility of exposure of the user or the analyzer to chemicals contained in the cartridge during the test and up to the disposal of used cartridges. Reagents may be found in three different physical forms: liquid, air-dried on surfaces or lyophilized powder cake. Within the cartridge, multiple steps are automatically performed in sequence by using pneumatic pressure and a multiport valve to transfer sample and fluids via the Transfer Chamber to their intended destinations.

QIAstat-Dx Analyzer 1.0: The QIAstat-Dx Analyzer 1.0 is the unit that hosts a cartridge and, on command from the user, is able to run predefined assay protocols. The software specific to this test is preloaded on the QIAstat-Dx Analyzer 1.0.

Other Materials: Each QIAstat-Dx Respiratory Panel cartridge will be used with a transfer pipette. The NPS sample from the patient will be collected in a sample tube using a swab in transport medium (not provided with device). QIAstat-Dx Analyzer - the QIAstat-Dx Respiratory Panel cartridge can only be run on the QIAstat-Dx Analyzer.

Calibrators and/or Controls: Blank controls are not applicable to the device because it is a single test disposable cartridge. Negative and positive external controls are recommended by the company but not provided with the QIAstat-Dx Respiratory Panel. QIAGEN provides an Internal Control within the QIAstat-Dx Respiratory Panel cartridge. The IC is an MS2 phage. The IC is located in the IC cavity and is mixed with the sample during sample preparation and the eluate is mixed with the Master Mix, then aliquoted in all Reaction Chambers. The primers and probes necessary to detect the IC are present in Reaction Chamber 1. The IC is a process control that will go through all nucleic acid extraction and amplification steps, similar to patient samples. The Analyzer 1.0 is provided factory calibrated and does not require user calibration. The Analyzer 1.0 includes self-check controls to verify the performance of all sensors and actuators and will alert the user in case of failure. The RCA will provide the results to the Application Software. The Application SW will store all the information related to a given result in the database and will display a summary of detected and equivocal analytes and the result for the IC. All POSITIVE or EQUIVOCAL analytes will be listed as "DETECTED PATHOGENS". The screen will also display the complete list of all "TESTED PATHOGENS", including positive, negative, equivocal or invalid analytes.

Specimen collection and transport materials: Samples are collected using a single-use Nasopharyngeal swab and a tube filled with transport medium. NPS swab specimens are to be collected and eluted using one of the following compatible collection kits: Universal Transport Medium (UTM™) (Copan Diagnostics (Brescia, Italy and CA, USA)), MicroTest™ M4, M4RT, M5, M6 (ThermoFisher Scientific, MA, USA), BD Universal Viral Transport (UVT) System (Becton Dickinson, NJ, USA), Universal Transport Medium (UTM) System (HealthLink Inc., FL, USA), Universal Transport Medium (Diagnostic Hybrids, OH, USA), V-C-M Medium (Quest Diagnostics, NJ, USA) or UniTranz-RT® Universal Transport Media (Puritan Diagnostics, ME, USA) collection kits.

Accessories and requirements: To be used in combination with the QIAstat-Dx Analyzer. Transfer pipette (MS-253003) used with each QIAstat-Dx® Respiratory Panel cartridge.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

nasopharyngeal swabs (NPS)

Indicated Patient Age Range

Not Found

Intended User / Care Setting

clinical laboratories

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

A total of 2,304 residual NPS specimens (1994 prospective and 310 archived) were tested in this comparison study. Between December 2017 to April 2019, specimens were prospectively collected from all comers meeting the study inclusion criteria and immediately frozen for later testing by the study site as frozen prospective specimens (N=1,093). No frozen samples were distributed amongst sites. At time of testing, specimens were thawed and tested on both the QIAstat-Dx Respiratory Panel and comparator method.

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

A total of 1994 specimens were evaluated for all panel members in the prospective study.

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Performance Characteristics - Non-clinical Studies
Limit of Detection (LoD): The LoD for each QIAstat-Dx Respiratory Panel pathogen was assessed by analyzing serial dilutions of analytical samples prepared from high-titer stocks obtained from commercial suppliers (ZeptoMetrix and ATCC) or artificial samples for commercially unavailable target analytes. The LoD concentration was determined for a total of 51 pathogen strains. The LoD of the QIAstat-Dx Respiratory Panel was determined per analyte using selected strains representing individual pathogens that are possible to detect with the QIAstat-Dx Respiratory Panel. To confirm the established LoD concentration, the detection rate of all replicates must be >=95% (at least 19/20 replicates must generate a positive signal). At least three different cartridge lots and at least three different QIAstat-Dx Analyzers were used for LoD determination for every pathogen.

Analytical Reactivity (Inclusivity): Evaluated with a collection of 127 respiratory pathogen isolates/strains selected based on clinical relevance and temporal/geographical diversity. Wet testing was done with specified strains, each tested in triplicates with a 100% detection rate for concentrations listed.

Analytical Specificity (Cross-Reactivity and Exclusivity): Carried out by in silico analysis and in vitro testing to assess cross-reactivity and exclusivity. On-panel organisms were tested for intra-panel cross-reactivity, and off-panel organisms (clinically relevant, common skin flora, lab contaminants, or microorganisms from previous infections) were tested for exclusivity. Samples were prepared by spiking potential cross-reactive organisms into simulated nasopharyngeal swab sample matrix at least 10^5 TCID50/ml for viral targets and 10^6 CFU/ml for bacterial and fungal targets, representing levels approximately 800-1,000,000-fold higher than the LoD. Cross-reactivity with Bordetella holmesii and Bordetella bronchiseptica was observed and predicted by in silico analysis.

Interference: Effect of 30 potentially interfering substances on detectability was evaluated. Substances were added to contrived samples (mix of organisms at 5xLoD) at levels above expected concentrations in NPS specimens. Endogenous substances (whole blood, human genomic DNA, pathogens) and exogenous substances (antibiotics, nasal sprays, workflow contaminants) were tested. All pathogen-containing samples without spiked interferent generated positive signals. Negative signals were obtained for pathogens not present in the sample. No inhibition was observed except for nasal influenza vaccines (Fluenz Tetra and FluMist), which were predicted to be reactive; final dilution without observable interfering effect was 0.000001% v/v. Clinically relevant co-infections testing demonstrated that all targets can be detected with multiple pathogens present.

Specimen Stability: Verification that storage of NPS samples at specified conditions does not impact performance. Ten sample mixes were used, with pathogens spiked into HeLa in UTM combined samples at 5xLoD or 1xLoD. 10 replicates per storage condition and target were tested. Storage conditions included 0h at 15-25°C, 4h at 15-25°C, 72h at 2-8°C, and 30 days at -15 to -25°C. Results supported recommendations for storage: up to 4h at RT (15-25°C), up to 3 days in fridge (2-8°C), and up to 30 days frozen (-15 to -25°C).

Matrix Equivalency: Comparison of performance between analytical samples prepared in NPS simulated matrix and negative clinical NPS sample matrix, and combined samples versus single-spiked samples. 4 combined sample mixes prepared by spiking individual pathogens in true-negative clinical NPS sample matrix. 1xLoD concentration for at least one strain per pathogen was tested in 20 replicates in true-negative clinical NPS sample matrix. LoD in clinical NPS sample matrix was not equivalent to LoD in simulated matrix for all analytes, but analytical studies used simulated matrix (more challenging condition).

Reproducibility: Testing of contrived samples at three test sites, incorporating variation factors (sites, days, replicates, cartridge lots, operators, analyzers). Testing over 5 days with 4 replicates/day (20 replicates/target/concentration/site), minimum 2 analyzers/site, and at least 2 operators/day. 12 sample mixes prepared with at least 3 replicates/mix, spiked at 0.1xLoD, 1xLoD, or 3xLoD.

At 0.1xLoD: detection rate for 24 of 24 targets was =95%.
At 3xLoD: detection rate for 24 of 24 targets was >=95%.

Performance Characteristics - Clinical Studies
Study Type: Multi-center study conducted at six (6) geographically diverse study sites (five U.S. and one international).
Sample Size: A total of 2,304 residual NPS specimens (1994 prospective and 310 archived).
Comparator: FDA cleared multiplexed respiratory pathogen panel that matched all panel members.
Key Results: The QIAstat-Dx Respiratory Panel detected a total of 191 specimens with distinctive multiple organism detections (9.6% of all specimens) in the prospective study. A total of 1994 prospective clinical specimens were tested and analyzed. Of these, 95.88% (1912/1994) yielded valid results on the first attempt. Invalid or no result were obtained for 82 specimens (4.11%). 72 of the 82 initially failed specimens yielded valid results after a single retesting using a new cartridge/sample. The remaining 10 specimens failed on the second attempt.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

The QIAstat-Dx Respiratory Panel prospective performance data in positive percent and negative percent agreements against the comparator methods are presented by analyte in Table 5.11 (Page 44) of the provided document.

Selected Overall Results (Sensitivity/PPA and Specificity/NPA with 95% CI):

Adenovirus:
- Sensitivity/PPA: 95.6% (89.1 - 98.3)
- Specificity/NPA: 99.2% (98.6 - 99.5)
Coronavirus 229E:
- Sensitivity/PPA: 88.9% (56.5-98.0)
- Specificity/NPA: 100.0% (99.8-100.0)
Coronavirus HKU1:
- Sensitivity/PPA: 98.1% (89.9-99.7)
- Specificity/NPA: 99.6% (99.3-99.8)
Coronavirus NL63:
- Sensitivity/PPA: 85.1% (72.3-92.6)
- Specificity/NPA: 99.9% (99.6-100.0)
Coronavirus OC43:
- Sensitivity/PPA: 89.7% (73.6-96.4)
- Specificity/NPA: 99.8% (99.5-99.9)
Human Metapneumovirus:
- Sensitivity/PPA: 94.3% (88.6-97.2)
- Specificity/NPA: 99.7% (99.4-99.9)
Rhinovirus/Enterovirus:
- Sensitivity/PPA: 91.2% (87.4-93.9)
- Specificity/NPA: 97.9% (97.1-98.5)
Influenza A:
- Sensitivity/PPA: 99.2% (97.0-99.8)
- Specificity/NPA: 99.5% (99.0-99.7)
Influenza A H1:
- Sensitivity/PPA: 0.0% (0.0-79.3)
- Specificity/NPA: 100.0% (99.8-100.0)
Influenza A H1N1/pdm09:
- Sensitivity/PPA: 98.8% (93.3-99.8)
- Specificity/NPA: 99.7% (99.4-99.9)
Influenza A H3:
- Sensitivity/PPA: 99.4% (96.5-99.9)
- Specificity/NPA: 99.6% (99.2-99.8)
Influenza B:
- Sensitivity/PPA: 94.6% (89.2-97.3)
- Specificity/NPA: 99.9% (99.7-100.0)
Parainfluenza 1:
- Sensitivity/PPA: 94.1% (73.0-99.0)
- Specificity/NPA: 99.8% (99.6-99.9)
Parainfluenza 2:
- Sensitivity/PPA: 100.0% (34.2-100.0)
- Specificity/NPA: 100.0% (99.8-100.0)
Parainfluenza 3:
- Sensitivity/PPA: 98.2% (93.8-99.5)
- Specificity/NPA: 99.7% (99.4-99.9)
Parainfluenza 4:
- Sensitivity/PPA: 100.0% (43.8-100.0)
- Specificity/NPA: 99.9% (99.6-100.0)
Respiratory Syncytial Virus (RSV):
- Sensitivity/PPA: 96.3% (93.0-98.1)
- Specificity/NPA: 99.7% (99.3-99.9)
Bordetella pertussis:
- Sensitivity/PPA: 100.0% (43.8-100.0)
- Specificity/NPA: 99.7% (99.3-99.9)
Chlamydophila pneumoniae:
- Sensitivity/PPA: 100.0% (56.6-100.0)
- Specificity/NPA: 99.9% (99.7-100.0)
Mycoplasma pneumoniae:
- Sensitivity/PPA: 100.0% (83.2-100.0)
- Specificity/NPA: 99.7% (99.4-99.9)

Predicate Device(s):

K123620 FilmArray® Respiratory Panel

Reference Device(s):

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.

(a)
Identification. A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B;
(2) Influenza A subtype H1 and Influenza A subtype H3;
(3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B;
(4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus;
(5) Human Metapneumovirus;
(6) Rhinovirus; and
(7) Adenovirus.
(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;”
(2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and
(3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.

Summary

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left, there is a symbol of the Department of Health & Human Services. To the right of the symbol, there is the FDA logo in blue, with the words "U.S. FOOD & DRUG" stacked on top of the word "ADMINISTRATION".

QIAGEN GmbH Melissa Mahall Sr. Director Regulatory Affairs 19300 Germantown Road Germantown, Maryland 20874 May 18, 2019

Re: K183597

Trade/Device Name: QIAstat-Dx Respiratory Panel Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay Regulatory Class: Class II Product Code: OCC, OEM, OOU, OEP, OOI, OTG, OZX, OZY, OQW, OZZ Dated: April 9, 2019 Received: April 9, 2019

Dear Melissa Mahall:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's

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requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely.

Uwe Scherf, Ph.D. Director Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K183597

Device Name QIAstat-Dx Respiratory Panel

Indications for Use (Describe)

The QIAstat-Dx Respiratory Panel is a multiplexed nucleic acid test intended for use with QIAstat-Dx system for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) eluted in Universal Transport Media (UTM) obtained from individuals suspected of respiratory tract infections. The following organism types are identified using the QIAstat-Dx Respiratory Panel: Adenovirus, Coronavirus 229E, Coronavirus HKU1, Coronavirus NL63, Coronavirus OC43, Human Metapneumovirus A+B, Influenza A, Influenza A H1, Influenza A H1N1/pdm09, Influenza B, Parainfluenza Virus 1, Parainfluenza Virus 2, Parainfluenza Virus 4, Rhinovirus/Enterovirus, Respiratory Syncytial Virus A+B, Bordetella pertussis, Chlamydophila pneumoniae and Mycoplasma pneumoniae.

The detection and identification of specific viral and bacterial nucleic acids from individuals presenting with signs and symptoms of a respiratory infection aids in the diagnosis of respiratory infection with other clinical and epidemiological information. The results of this test should not be used as the sole basis for diagnosis, treatment or other management decisions. Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by the test or lower respiratory tract infection that is not detected by a nasopharyngeal swab specimen. Positive results do not rule out co-infection with other organisms: the agent(s) detected by the QIAstat-Dx Respiratory Panel may not be the definite cause of disease. Additional laboratory testing (e.g., bacterial and viral culture, immunofluorescence and radiography) may be necessary when evaluating a patient with possible respiratory tract infection.

Due to the small number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Bordetella pertussis and Parainfluenza Virus 1 were established primarily with retrospective clinical specimens. Performance characteristics for Chlamydophila pneumoniae, Parainfluenza Virus 2, Parainfluenza Virus 4, Influenza A subtype H1 and Coronavirus 229E were established primarily using contrived clinical specimens.

Due to the genetic similarity between Human Rhinovirus and Enterovirus, the QIAstat-Dx Respiratory Panel cannot reliably differentiate them. A positive QI Respiratory Panel Rhinovirus/Enterovirus result should be followed-up using an alternate method (e.g., cell culture or sequence analysis).

Performance characteristics for Influenza A were established when Influenza A H1N1-2009 and A H3 were the predominant Influenza A viruses in circulation. Performance of detecting Influenza A may vary if other Influenza A strains are circulating or a novel Influenza A virus emerges. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

Type of Use (Select one or both, as applicable)

Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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Page 1 of 43

510(k) SUMMARY

General Information

| Submitted by: | QIAGEN GmbH
QIAGEN Strasse 1
Hilden, Germany D-40724 |
|-----------------|-------------------------------------------------------------------------------------------------------|
| Contact Person: | Melissa Mahall
Senior Director Regulatory Affairs
19300 Germantown Road
Germantown, MD 20874 |
| | Phone: 301-944-7768
Email: melissa.mahall@qiagen.com |
| Date Prepared: | April 9, 2019 |
| Device Name: | QIAstat-Dx® Respiratory Panel |
| Trade Name: | QIAstat-Dx® Respiratory Panel |
| Common Name: | QIAstat-Dx® Respiratory Panel |
| Classification: | 866.3980 - Respiratory viral panel multiplex nucleic acid assay |
| Product Code: | OCC, OEM, OOU, OEP, OTG, OQW, OOI, OZZ, OZY, OZX |

Predicate Device

Manufacturer	Product Name	510(k) No.
BioFire Diagnostics, Inc.	FilmArray® Respiratory Panel (RP)	K123620

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Device Description

QIAstat-Dx® is based on single-test cartridges with pre-packaged reagents including both wet and dry chemistry to handle the sample preparation and detection steps for the presence of a range of selected analytes by PCR technology. After insertion of the sample, the QIAstat-Dx assay cartridge is processed by the QIAstat-Dx® Analyzer 1.0.

Principle of Operation

The QIAstat-Dx® Respiratory Panel is part of the QIAstat-Dx® system and works with the QIAstat-Dx® Analyzer 1.0.

The OIAstat-Dx® Respiratory Panel is intended to be used with liquid sample nasopharyngeal swabs (NPS).

Once the cartridge has been inserted into the instrument, the test starts automatically and runs for approximately 74 minutes. When the test is finished, the cartridge is removed by the user and discarded. The QIAstat-Dx® Analyzer 1.0 automatically interprets test results and displays a summary on the analyzer display screen. The results can be printed using a connected printer if needed. The detected analytes are displayed in red. All other tested but not detected analytes are listed in green. The analyzer will report if an error occurs during processing, in which case the test will fail and no results will be provided (screen will show "FAIL").

Resuspension of IC and Prot K

Following insertion of the cartridge, the IC and Prot K are resuspended with the buffer located in Reservoir 1 (resuspension buffer from R1 is added to the interconnected IC cavity and Prot K cavity and transferred repeatedly between the Transfer Chamber and the cavities to ensure resuspension. The resuspended IC and Prot K are transferred to the sample cavity.

Cell Lysis

Primary lysis of the cells and analytes present in a NPS sample and IC occurs by a combination of chemical and mechanical processes using a rotor inside the lysis chamber in the presence of a buffer that acts as a chemical agent in aiding the mechanical process. The fast movement of the rotor results in sample agitation, which creates turbulence and shear forces that favor the lysis of the cell wall.

After mechanical lysis is completed, the primary lysate is transferred to the purification chamber through a frit with 80 um pore size. The second lysis buffer (from Reservoir 2) is added to the primary lysate to complete chemical lysis.

6

Purification

Binding reagent (from Reservoir 4) is added to the lysate in the purification cavity, and the mix is passed through the silica membrane. In this process, the DNA/RNA molecules stick to the membrane, and the remaining components of the lysate are delivered to the waste chamber. Then the membrane is washed with a first washing buffer (from Reservoir 5) to wash away proteins. This is followed by a second washing step with a second washing buffer (from Reservoir 6) to remove any remaining substances other than the nucleic acids. A subsequent drying step eliminates volatile substances from the silica membrane. Prior to the elution step, the Transfer Chamber is rinsed with the rinsing buffer (from Reservoir 7) in order to remove any potential inhibitors from previous processing steps. At the end of the process, the nucleic acids are released from the membrane using an elution buffer (from Reservoir 8). The eluate is collected in the Transfer Chamber.

Rehydration of Master Mix

A defined volume (approximately 135uL) of the eluate is delivered to the dry chemistry reservoir (DCC) to rehydrate the Master Mix. Any remaining eluate is transferred to the waste chamber. The eluate/Master Mix solution is mixed by repeated transfer between the Transfer Chamber and the DCC.

Aliquotting and PCR

Defined aliquots (approximately 15 uL) of mixed eluate/Master Mix are sequentially transferred from the Transfer Chamber to each of seven Reaction Chambers containing the specified, air dried primers and probes.

Within each Reaction Chamber, multiplex rtPCR testing is performed. Increase in fluorescence (indicative of detection of each target analyte) is detected directly within each Reaction Chamber.

The rtPCR process is conducted by two submodules of the QIAstat-Dx® Analyzer 1.0: the Thermal Cycler and the qPCR Sensor.

Components Description

QIAstat-Dx® Respiratory Panel Cartridge:

The QIAstat-Dx® Respiratory Panel cartridge is a disposable plastic device that allows performing fully automated molecular assays. The main features of the QIAstat-Dx® Respiratory Panel cartridge for the RP assay include the ability to test liquid samples as well as direct swabs and the capacity to store all necessary reagents within the cartridge needed for such testing. The cartridge is also designed to allow future expansion to incorporate additional sample types, such as swabs. All sample preparation and assay steps will be performed within the cartridge.

All the reagents required for the complete execution of the test are pre-loaded and selfcontained in the QIAstat-Dx® Respiratory Panel. The user does not need to manipulate

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any reagents. During the test, reagents are handled by pneumatically-operated microfluidics without any direct contact with the user or the analyzer actuators. This eliminates any possibility of exposure of the user or the analyzer to chemicals contained in the cartridge during the test and up to the disposal of used cartridges.

Reagents may be found in three different physical forms: liquid, air-dried on surfaces or lyophilized powder cake.

Within the cartridge, multiple steps are automatically performed in sequence by using pneumatic pressure and a multiport valve to transfer sample and fluids via the Transfer Chamber to their intended destinations.

QIAstat-Dx Analyzer 1.0

The QIAstat-Dx® Analyzer 1.0 is the unit that hosts a cartridge and, on command from the user, is able to run predefined assay protocols. The software specific to this test is preloaded on the QIAstat-Dx® Analyzer 1.0.

Other Materials

Each QIAstat-Dx® Respiratory Panel cartridge will be used with a transfer pipette. The NPS sample from the patient will be collected in a sample tube using a swab in transport medium (not provided with device).

QIAstat-Dx® Analyzer - the QIAstat-Dx® Respiratory Panel cartridge can only be run on the QIAstat-Dx® Analyzer.

Calibrators and/or Controls

Blank controls are not applicable to the device because it is a single test disposable cartridge. Negative and positive external controls are recommended by the company but not provided with the QIAstat-Dx® Respiratory Panel.

OIAGEN provides an Internal Control within the OIAstat-Dx® Respiratory Panel cartridge. The IC is an MS2 phage. The IC is located in the IC cavity and is mixed with the sample during sample preparation and the eluate is mixed with the Master Mix, then aliquoted in all Reaction Chambers. The primers and probes necessary to detect the IC are present in Reaction Chamber 1. The IC is a process control that will go through all nucleic acid extraction and amplification steps, similar to patient samples.

The Analyzer 1.0 is provided factory calibrated and does not require user calibration. The Analyzer 1.0 includes self-check controls to verify the performance of all sensors and actuators and will alert the user in case of failure.

The RCA will provide the results to the Application Software. The Application SW will store all the information related to a given result in the database and will display a summary of detected and equivocal analytes and the result for the IC. All POSITIVE or EQUIVOCAL analytes will be listed as "DETECTED PATHOGENS". The screen will

8

also display the complete list of all "TESTED PATHOGENS", including positive, negative, equivocal or invalid analytes.

Specimen collection and transport materials

Samples are collected using a single-use Nasopharyngeal swab and a tube filled with transport medium.

NPS swab specimens are to be collected and eluted using one of the following compatible collection kits: Universal Transport Medium (UTM™) (Copan Diagnostics (Brescia, Italy and CA, USA)), MicroTest™ M4, M4RT, M5, M6 (ThermoFisher Scientific, MA, USA), BD Universal Viral Transport (UVT) System (Becton Dickinson, NJ, USA), Universal Transport Medium (UTM) System (HealthLink Inc., FL, USA), Universal Transport Medium (Diagnostic Hybrids, OH, USA), V-C-M Medium (Quest Diagnostics, NJ, USA) or UniTranz-RT® Universal Transport Media (Puritan Diagnostics, ME , USA) collection kits.

Accessories and requirements

To be used in combination with the QIAstat-Dx Analyzer.

Transfer pipette (MS-253003) used with each QIAstat-Dx® Respiratory Panel cartridge.

Intended Use

The QIAstat-Dx® Respiratory Panel is a multiplexed nucleic acid test intended for use with QIAstat-Dx® system for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) eluted in Universal Transport Media (UTM) obtained from individuals suspected of respiratory tract infections. The following organism types and subtypes are identified using the QIAstat-Dx Respiratory Panel: Adenovirus, Coronavirus 229E, Coronavirus HKU1, Coronavirus NL63, Coronavirus OC43, Human Metapneumovirus A+B, Influenza A. Influenza A H1, Influenza A H3, Influenza A H1N1/pdm09, Influenza B, Parainfluenza Virus 1, Parainfluenza Virus 2, Parainfluenza Virus 3, Parainfluenza Virus 4, Rhinovirus/Enterovirus, Respiratory Syncytial Virus A+B, Bordetella pertussis, Chlamydophila pneumoniae, and Mycoplasma pneumoniae.

The detection and identification of specific viral and bacterial nucleic acids from individuals presenting with signs and symptoms of a respiratory infection aids in the diagnosis of respiratory infection if used in conjunction with other clinical and epidemiological information. The results of this test should not be used as the sole basis for diagnosis, treatment, or other management decisions. Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by the test or lower respiratory tract infection that is not detected by a nasopharyngeal swab specimen. Positive results do not rule out co-infection with other organisms: the agent(s) detected by the OIAstat-Dx Respiratory Panel may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and

9

radiography) may be necessary when evaluating a patient with possible respiratory tract infection.

Due to the small number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Bordetella pertussis and Parainfluenza Virus 1 were established primarily with retrospective clinical specimens. Performance characteristics for Chlamydophila pneumoniae, Parainfluenza Virus 2, Parainfluenza Virus 4, Influenza A subtype H1 and Coronavirus 229E were established primarily using contrived clinical specimens.

Due to the genetic similarity between Human Rhinovirus and Enterovirus, the QIAstat-Dx Respiratory Panel cannot reliably differentiate them. A positive OIAstat-Dx Respiratory Panel Rhinovirus/Enterovirus result should be followed-up using an alternate method (e.g., cell culture or sequence analysis).

Performance characteristics for Influenza A were established when Influenza A H1N1-2009 and A H3 were the predominant Influenza A viruses in circulation. Performance of detecting Influenza A may vary if other Influenza A strains are circulating or a novel Influenza A virus emerges. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

Comparison of the OIAstat-Dx® Respiratory Panel and the Predicate Device

The QIAstat-Dx® Respiratory Panel is substantially equivalent to the predicate device:

K123620: FilmArray® Respiratory Panel
Similarities and differences between the QIAstat-Dx® Respiratory Panel and the predicate device are shown in Table 5.1.

Characteristic	Device	Predicate
Name	QIAstat-Dx® Respiratory Panel	BioFire Diagnostics, Inc.'s
FilmArray® Respiratory Panel
(RP)


510(k) No.	K183597	K123620
Regulation	21 CFR 866.3980	21 CFR 866.3980
Product Code	OCC	OCC
Device Class	Class II	Class II
Characteristic	Device	Predicate
	Similarities
Intended Use	The QIAstat-Dx Respiratory Panel
is a multiplexed nucleic acid test
intended for use with QIAstat-Dx
system for the simultaneous
qualitative detection and
identification of multiple
respiratory viral and bacterial
nucleic acids in nasopharyngeal
swabs (NPS) eluted in universal
transport media (UTM) obtained
from individuals suspected of
respiratory tract infections. The
following organism types and
subtypes are identified using the
QIAstat-Dx Respiratory Panel:
Adenovirus, Coronavirus 229E,
Coronavirus HKU1, Coronavirus
NL63, Coronavirus OC43, Human
Metapneumovirus A+B, Influenza
A, Influenza A H1, Influenza A
H3, Influenza A H1N1/pdm09,
Influenza B, Parainfluenza Virus
1, Parainfluenza Virus 2,
Parainfluenza Virus 3,
Parainfluenza Virus 4,
Rhinovirus/Enterovirus,
Respiratory Syncytial Virus A+B,
Bordetella pertussis,
Chlamydophila pneumoniae, and
Mycoplasma pneumoniae.	FilmArray® Respiratory Panel
(RP) is a multiplexed nucleic acid
test intended for use with the
FilmArray Instrument for the
simultaneous qualitative detection
and identification of multiple
respiratory viral and bacterial
nucleic acids in nasopharyngeal
swabs (NPS) obtained from
individuals suspected of
respiratory tract infections. The
following organism types and
subtypes are identified using the
FilmArray RP: Adenovirus,
Coronavirus 229E, Coronavirus
HKU1, Coronavirus NL63,
Coronavirus OC43, Human
Metapneumovirus, Influenza A,
Influenza A subtype H1,
Influenza A subtype H3,
Influenza A subtype H1-2009,
Influenza B, Parainfluenza Virus
1, Parainfluenza Virus 2,
Parainfluenza Virus 3,
Parainfluenza Virus 4, Human
Rhinovirus/Enterovirus,
Respiratory Syncytial Virus,
Bordetella pertussis,
Chlamydophila pneumoniae, and
Mycoplasma pneumoniae. The
detection and identification of
specific viral and bacterial nucleic
acids from individuals exhibiting
signs and symptoms of a
respiratory infection aids in the
diagnosis of respiratory infection
if used in conjunction with other
clinical and epidemiological
information. The results of this
test should not be used as the sole
basis for diagnosis, treatment, or
other management decisions.
	The detection and identification of
specific viral and bacterial nucleic
acids from individuals presenting
with signs and symptoms of a
respiratory infection aids in the
diagnosis of respiratory infection
if used in conjunction with other
clinical and epidemiological
information. The results of this test
should not be used as the sole
basis for diagnosis, treatment, or
other management decisions.	Negative results in the setting of a
Characteristic	Device	Predicate
	Negative results in the setting of a
respiratory illness may be due to
infection with pathogens that are
not detected by the test or lower
respiratory tract infection that is
not detected by a nasopharyngeal
swab specimen. Positive results do
not rule out co-infection with other
organisms: the agent(s) detected
by the QIAstat-Dx Respiratory
Panel may not be the definite
cause of disease. Additional
laboratory testing (e.g. bacterial
and viral culture,
immunofluorescence, and
radiography) may be necessary
when evaluating a patient with
possible respiratory tract infection.	respiratory illness may be due to
infection with pathogens that are
not detected by this test or, lower
respiratory tract infection that is
not detected by a nasopharyngeal
swab specimen. Positive results
do not rule out coinfection with
other organisms: the agent(s)
detected by the Film Array RP
may not be the definite cause of
disease. Additional laboratory
testing (e.g. bacterial and viral
culture, immunofluorescence, and
radiography) may be necessary
when evaluating a patient with
possible respiratory tract
infection.
	Due to the small number of
positive specimens collected for
certain organisms during the
prospective clinical study,
performance characteristics for
Bordetella pertussis and
Parainfluenza Virus 1 were
established primarily with
retrospective clinical specimens.	Due to the small number of
positive specimens collected for
certain organisms during the
prospective clinical study,
performance characteristics for
Bordetella pertussis, Coronavirus
229E, Coronavirus OC43,
Influenza A H1, Influenza A H3,
Influenza A H1-2009, Influenza
B, Mycoplasma pneumoniae,
Parainfluenza Virus 1,
Parainfluenza Virus 2, and
Parainfluenza Virus 4 were
established primarily with
retrospective clinical specimens.
	Performance characteristics for
Chlamydophila pneumoniae,
Parainfluenza Virus 2,
Parainfluenza Virus 4, Influenza A
subtype H1 and Coronavirus 229E
were established primarily using
contrived clinical specimens.	Performance characteristics for
Chlamydophila pneumoniae were
established primarily using
contrived clinical specimens.
	Due to the genetic similarity
between Human Rhinovirus and
Enterovirus, the QIAstat-Dx
Respiratory Panel cannot reliably
differentiate them. A positive
QIAstat-Dx Respiratory Panel
Rhinovirus/Enterovirus result
should be followed up using an	Due to the genetic similarity
between Human Rhinovirus and
Enterovirus, the FilmArray RP
cannot reliably differentiate them.
A positive FilmArray RP
Rhinovirus/Enterovirus result
should be followed-up using an
alternate method (e.g., cell culture
Characteristic	Device	Predicate
	alternate method (e.g., cell culture
or sequence analysis).
Performance characteristics for
Influenza A were established when
Influenza A H1N1-2009 and A H3
were the predominant Influenza A
viruses in circulation. Performance
of detecting Influenza A may vary
if other Influenza A strains are
circulating or a novel Influenza A
virus emerges. If infection with a
novel Influenza A virus is
suspected based on current clinical
and epidemiological screening
criteria recommended by public
health authorities, specimens
should be collected with
appropriate infection control
precautions for novel virulent
Influenza viruses and sent to state
or local health departments for
testing. Viral culture should not be
attempted in these cases unless a
BSL 3+ facility is available to
receive and culture specimens.	or sequence analysis).
The FilmArray RP assay for
Coronavirus OC43 may cross-
react with some isolates of
Coronavirus HKU1. A dual
positive result may be due to
cross-reactivity or may indicate a
co-infection.
Performance characteristics for
Influenza A were established
when Influenza A H1-2009, A
H1, and A H3 were the
predominant Influenza A viruses
in circulation. Performance of
detecting Influenza A may vary if
other Influenza A strains are
circulating or a novel Influenza A
virus emerges. If infection with a
novel Influenza A virus is
suspected based on current
clinical and epidemiological
screening criteria recommended
by public health authorities,
specimens should be collected
with appropriate infection control
precautions for novel virulent
Influenza viruses and sent to state
or local health departments for
testing. Viral culture should not
be attempted in these cases unless
a BSL 3+ facility is available to
receive and culture specimens.
Specimen Type	Nasopharyngeal swabs (NPS)
eluted in UTM	Nasopharyngeal swabs (NPS)
Assay Targets
Amplification
and Detection
Technology	See analyte list above, RNA/ DNA
PCR	See analyte list above, RNA/DNA
PCR
Assay Controls	One internal control in each
cartridge to control for sample
processing that is subjected to all
nucleic acid extraction and	Two controls are included in each
reagent pouch to control for
sample processing and both stages
of PCR and melt analysis.
Characteristic	Device	Predicate
	amplification steps similar to
patient samples. Labeling will
recommend use of negative and
positive external controls
regularly. Use transport medium
as the external Negative Control,
and previously characterized
positive samples or negative
sample spiked with well
characterized target organisms as
external Positive Controls.	Labeling recommends the use of
external positive and negative
controls regularly. Use viral
transport medium as the external
negative control, and previously
characterized positive samples or
negative samples spiked with well
characterized organisms as
external positive controls.
	Differences
Nucleic Acid
Extraction	Extraction of nucleic acids using
spin columns	Extraction of nucleic acids using
magnetic beads
Amplification
and Detection
Instrument
System	QIAstat-Dx Analyzer	FilmArray Instrument

Table 5.1: Comparison of the QIAstat-Dx® Respiratory Panel with the predicate device

10

11

12

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Performance Characteristics - Non-clinical Studies

Limit of Detection

The Limit of Detection (LoD) is defined as the lowest concentration at which ≥95% of the tested samples generate a positive call. The LoD for each QIAstat-Dx Respiratory Panel pathogen was assessed by analyzing serial dilutions of analytical samples prepared from high-titer stocks obtained from commercial suppliers (ZeptoMetrix and ATCC) or artificial samples for commercially unavailable target analytes.

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

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

Individual LoD values for each QIAstat-Dx Respiratory Panel target is shown in Table 5.2.

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| Pathogen | Strain | Source | Concentration | Detection
rate |
|------------------------------------|--------------------------------------|-----------------------------|---------------------|-----------------------------|
| Influenza A H1N1 | A/New Jersey/8/76 | ATCC® VR-897 | 341 CEID50/ml | Flu A: 20/20
H1: 20/20 |
| | A/Brisbane/59/07 | ZeptoMetrix®
0810244CFHI | 4 TCID50/ml | Flu A: 20/20
H1: 20/20 |
| | A/New Caledonia/20/99 | ZeptoMetrix
0810036CFHI | 15 TCID50/ml | Flu A: 20/20
H1: 19/20 |
| Influenza A H3N2 | A/Virginia/ATCC6/2012 * | ATCC VR-1811 | 0.1 PFU/ml | Flu A: 20/20
H3: 20/20 |
| | A/Wisconsin/67/2005 * | ZeptoMetrix
0810252CFHI | 3.8 TCID50/ml | Flu A: 20/20
H3: 20/20 |
| | A/Port Chalmers/1/73 | ATCC VR-810 | 499.3
CEID50/ml | Flu A: 20/20
H3: 20/20 |
| Influenza A,
subtype H1N1/2009 | A/Virginia/ATCC1/2009 | ATCC VR-1736 | 67 PFU/ml | Flu A: 20/20
H1N1: 20/20 |
| | A/SwineNY/03/2009 | ZeptoMetrix
0810249CFHI | 56 TCID50/ml | Flu A: 20/20
H1N1: 20/20 |
| Influenza B | B/Virginia/ATCC5/2012 * | ATCC VR-1807 | 0.03 PFU/ml | 20/20 |
| | B/FL/04/06 | ATCC VR-1804 | 1080
CEID50/ml | 20/20 |
| | B/Taiwan/2/62 | ATCC VR-295 | 5000
CEID50/ml | 19/20 |
| Coronavirus 229E | — | ATCC VR-740 | 0.2 TCID50/ml | 20/20 |
| | — * | ZeptoMetrix
0810229CFHI | 3.6 TCID50/ml | 20/20 |
| Coronavirus OC43 | — | ATCC VR-1558 | 0.1 TCID50/ml | 20/20 |
| | — * | ZeptoMetrix
0810024CFHI | 0.1 TCID50/ml | 20/20 |
| Coronavirus NL63 | — | ZeptoMetrix
0810228CFHI | 0.01
TCID50/ml | 20/20 |
| Coronavirus HKU1 | — * | Clinical Sample
S510 | 40,000
copies/ml | 20/20 |
| Parainfluenza Virus
1 (PIV 1) | C35 * | ATCC VR-94 | 0.2 TCID50/ml | 19/20 |
| | — | ZeptoMetrix
0810014CFHI | 0.2 TCID50/ml | 19/20 |
| Pathogen | Strain | Source | Concentration | Detection rate |
| Parainfluenza Virus
2 (PIV 2) | Greer | ATCC VR-92 | 7.3 TCID50/ml | 20/20 |
| Parainfluenza Virus
2 (PIV 2) | - * | ZeptoMetrix
0810015CFHI | 1.3 TCID50/ml | 19/20 |
| Parainfluenza Virus
3 (PIV 3) | C 243 | ATCC VR-93 | 2.3 TCID50/ml | 20/20 |
| Parainfluenza Virus
3 (PIV 3) | - * | ZeptoMetrix
0810016CFHI | 11.5 TCID50/ml | 20/20 |
| Parainfluenza Virus
4a (PIV 4a) | M-25 | ATCC VR-1378 | 0.5 TCID50/ml | 20/20 |
| Parainfluenza Virus
4b (PIV 4b) | - * | ZeptoMetrix
0810060BCFHI | 9.5 TCID50/ml | 20/20 |
| Respiratory
Syncytial Virus A | A2 * | ATCC VR-1540 | 12.0 PFU/ml | 20/20 |
| Respiratory
Syncytial Virus A | Long * | ATCC VR-26 | 33.0 PFU/ml | 20/20 |
| Respiratory
Syncytial Virus B | 18537 * | ATCC VR-1580 | 0.03 PFU/ml | 20/20 |
| Respiratory
Syncytial Virus B | CH93(18)-18 | ZeptoMetrix
0810040CFHI | 0.4 TCID50/ml | 20/20 |
| Human
Metapneumovirus | Peru6-2003 (type B2) * | ZeptoMetrix
0810159CFHI | 0.01
TCID50/ml | 19/20 |
| Human
Metapneumovirus | hMPV-16, IA10-2003
(A1) | ZeptoMetrix
0810161CFHI | 0.5 TCID50/ml | 20/20 |
| Human
Metapneumovirus | hMPV-20, IA14-2003
(A2) * | ZeptoMetrix,
0810163CFHI | 0.4 TCID50/ml | 19/20 |
| Human
Metapneumovirus | hMPV-3, Peru2-2002
(B1) * | ZeptoMetrix,
0810156CFHI | 1479.9
TCID50/ml | 19/20 |
| Adenovirus | GB (Adenovirus B3) | ATCC VR-3 | 4993.0
TCID50/ml | 20/20 |
| Adenovirus | RI-67 (Adenovirus E4) * | ATCC VR-1572 | 15.8
TCID50/ml | 20/20 |
| Adenovirus | Adenoid 75 (Adenovirus
C5) * | ATCC VR-5 | 7331.0
TCID50/ml | 20/20 |
| Adenovirus | Adenoid 71 (Adenovirus
C1) * | ATCC VR-1 | 69.5
TCID50/ml | 20/20 |
| Adenovirus | Adenoid 6 (Adenovirus
C2) * | ATCC VR-846 | 28.1
TCID50/ml | 20/20 |
| Adenovirus | Tonsil 99 (Adenovirus
C6) * | ATCC VR-6 | 88.8
TCID50/ml | 20/20 |
| Pathogen | Strain | Source | Concentration | Detection
rate |
| Enterovirus | /US/IL/14-18952
(Enterovirus D68) | ATCC VR-
1824 | 8.9
TCID50/ml | 19/20 |
| | Echovirus 6 * | ATCC VR-241 | 0.9
TCID50/ml | 19/20 |
| Rhinovirus | 1059 (Rhinovirus
B14) * | ATCC VR-284 | 8.9
TCID50/ml | 20/20 |
| | HGP (Rhinovirus A2) | ATCC VR-482 | 8.9
TCID50/ml | 19/20 |
| | 11757 (Rhinovirus
C16) * | ATCC VR-283 | 50.0
TCID50/ml | 20/20 |
| | Type 1A * | ATCC VR-
1559 | 8.9
TCID50/ml | 20/20 |
| Mycoplasma
pneumoniae | M129-B7 (type 1) * | ATCC 29342 | 0.1 CCU/ml | 20/20 |
| | PI 1428 | ATCC 29085 | 1.0 CCU/ml | 20/20 |
| Chlamydia
pneumoniae | TW183 | ATCC VR-
2282 | 14.2 IFU/ml | 20/20 |
| | CWL-029 * | ATCC VR-
1310 | 120.0 IFU/ml | 19/20 |
| Bordetella
pertussis | I028 | ATCC BAA-
2707 | 0.3 CFU/ml | 20/20 |
| | 18323 * | ATCC 9797 | 2.6 CFU/ml | 19/20 |

Table 5.2: LoD values obtained for the different respiratory target strains tested with the QIAstat-Dx Respiratory Panel

15

16

NOTE: For pathogen strains with (*), the LoD has been obtained in simulated matrix.

Analytical Reactivity

Analytical reactivity (Inclusivity) was evaluated with a collection of 127 respiratory pathogen isolates/strains that were selected based on clinical relevance and temporal/geographical diversity. Based on wet testing and in silico analysis, the QIAstat-Dx® Respiratory Panel primers and probes are specific and inclusive for clinically prevalent and relevant strains for each pathogen. Wet testing has been done with the strains listed in Table 5.3. Every strain has been tested in triplicates with a 100% detection rate for concentrations listed.

17

Pathogen	Strain	Supplier	Catalogue ID	Concentration tested	x-fold LoD
Influenza H1N1	A/Brisbane/59/07 a	Zeptometrix	0810244CF HI	0.4 TCID50/mL	1x LoD
	A/New Caledonia/20/99	Zeptometrix	0810036CF HI	1.5 TCID50/mL	0.3x LoD
	A/New Jersey/8/76	ATCC	VR-897	34.1 CEID50/mL	1x LoD
	A/Denver/1/57	ATCC	VR-546	340 CEID50/mL	0.1x LoD
	A/Mal/302/54	ATCC	VR-98	15.8 CEID50/mL	1x LoD
	A/Weiss/43	ATCC	VR-96	28117.1 CEID50/mL	0.1x LoD
	A/PR/8/34	ATCC	VR-1469	390 PFU/mL	3x LoD
	A/Fort Monmouth/1/1947	ATCC	VR-1754	28.1 CEID50/mL	0.1x LoD
	A/WS/33	ATCC	VR-1520	15.8 TCID50/mL	0.1x LoD
	A/Swine/Iowa/15/1930	ATCC	VR-333	889.1 CEID50/mL	1x LoD
Influenza H3N2	A/Port Chalmers/1/73 a	ATCC	VR-810	499.3 CEID50/mL	1x LoD
	A/Virginia/ATCC6/2012	ATCC	AV-VR-1811	0.1 PFU/mL	1x LoD
	A/Wisconsin/67/2005	Zeptometrix	0810252CF HI	3.8 TCID50/mL	1x LoD
	A/Wisconsin/15/2009	ATCC	VR-1882	5.8 CEID50/mL	1x LoD
	A/Victoria/3/75	ATCC	VR-822	16 CEID50/mL	1x LoD
	A/Aichi/2/68	ATCC	VR-1680	31 PFU/mL	10x LoD
	A/Hong Kong/8/68	ATCC	VR-1679	1581.1 TCID50/mL	10x LoD
	A/Alice (recombinant, carries A/England/42/72)	ATCC	VR-776	500 TCID50/mL	10x LoD
Pathogen	Strain	Supplier	Catalogue
ID	Concentration
tested	x-fold
LoD
	MRC-2 (recombinant
A/England/42/72 and
A/PR/8/34 strains)	ATCC	VR-777	8891.4
CEID50/mL	100x
LoD
	A/Switzerland/9715293/
2013	ATCC	VR-1837	1000
CEID50/mL	1x LoD
	A/Virginia/ATCC1/2009a	ATCC	VR-1736	6.7 PFU/mL	1x LoD
	A/SwineNY/03/2009	Zeptometrix	0810249CF
HI	5.6 TCID50/mL	1x LoD
	A/Virginia/ATCC2/2009	ATCC	VR-1737	61 PFU/mL	0.1x LoD
	A/Virginia/ATCC3/2009	ATCC	VR-1738	1800 PFU/mL	100x
LoD
Influenza A
H1N1 pan	Swine NY/01/2009	Zeptometrix	0810248CF
HI	138 TCID50/mL	0.3x LoD
	Swine NY/02/2009	Zeptometrix	0810109CF
NHI	1.4 TCID50/mL	10x LoD
	A/California/07/2009
NYMC X-179A	ATCC	VR-1884	1400
CEID50/mL	0.1x LoD
	Canada/6294/09	Zeptometrix	0810109CF
JHI	1.7 TCID50/mL	3x LoD
	Mexico/4108/09	Zeptometrix	0810166CF
HI	14.1
TCID50/mL	0.1x LoD
	Netherlands/2629/2009	BEI
Resources	NR-19823	16 TCID50/mL	0.3x LoD
Influenza A
H2N2	Japan/305/1957 (nucleic
acid)b	BEI	NR-2775	0.00326 RNA
ng/µL	1x LoD
	Korea/426/1968xPuerto
Rico/8/1934 (nucleic
acid)b	BEI	NR-9679	0.0000625
RNA ng/µL	0.3x LoD
Influenza A
H5N3	A/Duck/Singapore/645/1
997 (nucleic acid)b	BEI	NR-9682	0.002475 RNA
ng/µL	1x LoD
Influenza A
H10N7	Chicken/Germany/N/49
(nucleic acid)b	BEI	NR-2765	0.068 RNA
ng/µL	10x LoD
Pathogen	Strain	Supplier	Catalogue
ID	Concentration
tested	x-fold
LoD
Influenza A
H1N2	Recombinant Kilbourne
F63, A/NWS/1934 (HA)
x A/Rockefeller
Institute/5/1957 (nucleic
acids) b	BEI	NR-9677	0.0148 RNA
ng/µL	100x
LoD
	B/Virginia/ATCC5/2012 a	ATCC	VR-1807	0.03 PFU/mL	1x LoD
	B/FL/04/06	ATCC	VR-1804	108 CEID50/mL	1x LoD
Influenza B	B/Taiwan/2/62	ATCC	VR-295	49.9 CEID50/mL	0.3x LoD
	B/Allen/45 c	ATCC	VR-102	n/a	Not
detected
	B/Hong Kong/5/72 c	ATCC	VR-823	n/a	Not
detected
	B/Maryland/1/59	ATCC	VR-296	338 CEID50/mL	0.1x LoD
	B/GL/1739/54	ATCC	VR-103	50 CEID50/mL	1x LoD
	B/Wisconsin/1/2010	ATCC	VR-1883	0.3 CEID50/mL	0.1x LoD
	B/Massachusetts/2/2012	ATCC	VR-1813	2300
CEID50/mL	3x LoD
	B/Florida/02/06 d	Zeptometrix	0810037CF
HI	n/a	n/a
	B/Brisbane/60/2008	BEI
Resources	NR-42005	1.8 CEID50/mL	0.1x LoD
	B/Malaysia/2506/2004	BEI
Resources	NR-9723	1.58
CEID50/mL	0.3x LoD
Coronavirus
229E	n/a a	Zeptometrix	0810229CF
HI	3.6 TCID50/mL	1x LoD
	n/a	ATCC	VR-740	0.2 TCID50/mL	0.3x LoD
Coronavirus
OC43	n/a a	ATCC	VR-1558	0.1 TCID50/mL	1x LoD
	n/a	Zeptometrix	0810024CF
HI	0.1 TCID50/mL	1x LoD
Coronavirus
NL63	n/a a	Zeptometrix	0810228CF
HI	0.01
TCID50/mL	1x LoD
	n/a	BEI
Resources	NR-470	1.6 TCID50/mL	1x LoD
Pathogen	Strain	Supplier	Catalogue
ID	Concentration
tested	x-fold
LoD
Coronavirus
HKU1	n/a a, e	Zeptometrix	NATRVP-
IDI	3E+03
copies/mL	1x LoD
	n/a e	QIAGEN
Barcelona
(STAT-Dx)	Clinical
sample
S510	1.2E+04
copies/mL	0.3x LoD
	n/a e	QIAGEN
Barcelona
(STAT-Dx)	Clinical
sample
S501	7E+03
copies/mL	1x LoD
	n/a e	QIAGEN
Barcelona
(STAT-Dx)	Clinical
sample
S496	7E+03
copies/mL	1x LoD
Parainfluenza
Virus 1	n/a a	Zeptometrix	0810014CF
HI	0.02
TCID50/mL	1x LoD
	C35	ATCC	VR-94	0.2 TCID50/mL	1x LoD
	n/a	Zeptometrix	NATRVP-
IDI	1.0E-2 f	10x LoD
	Greer a	ATCC	VR-92	2.3 TCID50/mL	1x LoD
Parainfluenz
a Virus 2	n/a	Zeptometrix	0810015CF
HI	1.3 TCID50/mL	0.3x LoD
	n/a	Zeptometrix	0810504CF
HI	1.3 TCID50/mL	0.1x LoD
Parainfluenz
a Virus 3	n/a a	Zeptometrix	0810016CF
HI	11.5
TCID50/mL	1x LoD
	C 243	ATCC	VR-93	2.3 TCID50/mL	1x LoD
	n/a	Zeptometrix	NATRVP-
IDI	1.0E-3 f	0.1x LoD
Parainfluenz
a Virus 4	M-25 a	ATCC	VR-1378	0.5 TCID50/mL	1x LoD
	n/a	Zeptometrix	0810060B
CFHI	9.6 TCID50/mL	0.3x LoD
	n/a	Zeptometrix	0810060CF
HI	28.2
TCID50/mL	0.1x LoD
	CH 19503	ATCC	VR-1377	1 TCID50/mL	0.3x LoD
	18537 a	ATCC	VR-1580	0.03 PFU/mL	1x LoD
	A2	ATCC	VR-1540	12 PFU/mL	0.3x LoD
	Long	ATCC	VR-26	33 PFU/mL	1x LoD
Respiratory
Syncytial
Virus A+B	CH93(18)-18	Zeptometrix	0810040CF
HI	0.4 TCID50/mL	1x LoD
	n/a	Zeptometrix	0810040A
CFHI	0.3 TCID50/mL	0.1x LoD
	B WV/14617/85	ATCC	VR-1400	15.8
TCID50/mL	1x LoD
Human
Metapneumo
virus	IA10-2003 a	Zeptometrix	0810161CF
HI	0.5 TCID50/mL	1x LoD
	IA14-2003	Zeptometrix	0810163CF
HI	0.4 TCID50/mL	1x LoD
Pathogen	Strain	Supplier	Catalogue
ID	Concentration
tested	x-fold
LoD
	Peru2-2002	Zeptometrix	0810156CF
HI	1478.9
TCID50/mL	1x LoD
	Peru6-2003	Zeptometrix	0810159CF
HI	0.01
TCID50/mL	1x LoD
	IA3-2002	Zeptometrix	0810160CF
HI	66 TCID50/mL	3x LoD
	IA27-2004	Zeptometrix	0810164CF
HI	1.3 TCID50/mL	1x LoD
	Peru3-2003	Zeptometrix	0810158CF
HI	31.6
TCID50/mL	1x LoD
	IA18-2003	Zeptometrix	0810162CF
HI	0.4 TCID50/mL	1x LoD
	Peru1-2002	Zeptometrix	0810157CF
HI	2187.8
TCID50/mL	10x LoD
Adenovirus	Tonsil 99 a	ATCC	VR-6	88.8
TCID50/mL	1x LoD
	GB	ATCC	VR-3	4992.8
TCID50/mL	0.3x LoD
	Adenoid 71	ATCC	VR-1	69.5
TCID50/mL	1x LoD
	Adenoid 6	ATCC	VR-846	28.1
TCID50/mL	0.3x LoD
	Adenoid 75	ATCC	VR-5	7331.2
TCID50/mL	0.3x LoD
	RI-67	ATCC	VR-1572	15.8
TCID50/mL	0.3x LoD
	Huie	ATCC	VR-863	88.9
TCID50/mL	0.3x LoD
	Gomen	ATCC	VR-7	0.3 TCID50/mL	0.1x LoD
	Slobitski	ATCC	VR-12	16 TCID50/mL	10x LoD
	AV-1645 [128]	ATCC	VR-256	2.8 TCID50/mL	0.3x LoD
	Compton	ATCC	VR-716	0.28
TCID50/mL	0.3x LoD
Holden	ATCC	VR-718	8.9 TCID50/mL	0.3x LoD
Trim	ATCC	VR-1815	160 TCID50/mL	0.3x LoD
Dugan	ATCC	VR-931	0.2 TCID50/mL	0.1x LoD
Tak (73-3544)	ATCC	VR-930	28117
TCID50/mL	3x LoD
Enterovirus	/US/IL/14-18952 a	ATCC	VR-1824	8.9 TCID50/mL	1x LoD
	D-1 (Cox)	ATCC	VR-241	0.9 TCID50/mL	0.3x LoD
	H	ATCC	VR-1432	8.9 TCID50/mL	1x LoD
	M.K. (Kowalik)	ATCC	VR-168	1.0E-6 f	10x LoD
	Gregory	ATCC	VR-41	889.1
TCID50/mL	10x LoD
Pathogen	Strain	Supplier	Catalogue
ID	Concentration
tested	x-fold
LoD
	Bastianni	ATCC	VR-1660	281.2
TCID50/mL	1x LoD
	Griggs	ATCC	VR-1311	1.6 TCID50/mL	0.3x LoD
	Conn-5	ATCC	VR-28	158.1
TCID50/mL	0.3x LoD
	Ohio-1	ATCC	VR-29	2811.7
TCID50/mL	3x LoD
	Nancy	ATCC	VR-30	0.9 TCID50/mL	0.3x LoD
	CHHE-29	ATCC	VR-47	0.03
TCID50/mL	10x LoD
	Kuykendall [V-024-001-
012]	ATCC	VR-850	28.1
TCID50/mL	10x LoD
	1059 a	ATCC	VR-284	8.9 TCID50/mL	1x LoD
	2060	ATCC	VR-1559	8.9 TCID50/mL	0.1x LoD
	HGP	ATCC	VR-482	8.9 TCID50/mL	1x LoD
Rhinovirus	11757	ATCC	VR-283	49.9
TCID50/mL	0.3x LoD
	FEB	ATCC	VR-483	281.2
TCID50/mL	1x LoD
	33342	ATCC	VR-1663	200 PFU/mL	3x LoD
	PI 1428 a	ATCC	29085	1 CCU/mL	1x LoD
M. pneumoniae	M129-B7	ATCC	29342	0.1 CCU/mL	1x LoD
	FH strain of Eaton Agent
[NCTC 10119]	ATCC	15531	0.2 CFU/mL	0.1x LoD
B. pertussis	I028 a	ATCC	BAA-2707	0.3 CFU/mL	1x LoD
	19323	ATCC	9797	2.6 CFU/mL	1x LoD
	10-536	ATCC	10380	$1.0E-2$ f	0.3x LoD
C. pneumoniae	TW183 a	ATCC	VR-2282	14.2 IFU/mL	1x LoD
	CWL-029	ATCC	VR-1310	120 IFU/mL	1x LoD
	AR-39	ATCC	53592	29 IFU/mL	0.3x LoD

Table 5.3: In vitro Analytical Reactivity details for all the pathogens tested with the QIAstat-Dx® Respiratory Panel

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Page 19 of 43

ª LoD reference strain used to calculate the x-fold LoD.

b Influenza A/Brisbane/59/07 (Zeptometrix, 0810244CFHI) used as reference strain to calculate the x-fold LoD.

· In silico analysis showed that this strain should be detected by QIAstat-Dx Respiratory Panel V1. In in vitro testing, the strain was not detected. It is identified as a derivative from B/Lee/40 ancestral lineage which is not in circulation since the 1980s (Nogales A., Martínez-Sobrido L. (2017). Reverse Genetics Approaches for the Development of Influenza Vaccines (Review). Int. J. Mol. Sci. 2017, 18, 20.).

4 In silico analysis showed that this strain should be detected by QIAstat-Dx Respiratory Panel V1. In in vitro testing, the strain (Victoria lineage) was randomly detected, therefore x-fold LoD could not be determined.

& Coronavirus HKU1 was quantified by a real-time PCR assay against a standard curve of synthetic Coronavirus HKU1 RNA transcript to obtain quantification of the viral nucleic acid in the clinical specimen (RNA copies/mL).

f Relative dilution from stock. Stock titer not available according to manufacturer.

23

Analytical Specificity (Cross-Reactivity and Exclusivity)

The analytical specificity study was carried out by in silico analysis and in vitro testing to assess the cross-reactivity and exclusivity of the QIAstat-Dx Respiratory Panel. On-panel organisms were tested to assess the potential for intra-panel cross-reactivity and off-panel organisms were tested to evaluate panel exclusivity. The off-panel organisms selected were clinically relevant organisms (colonizing the upper respiratory tract or causing respiratory symptoms), common skin flora or laboratory contaminants, or microorganisms for which much of the population may have been infected. The on-panel organisms tested are shown in Table 5.4.

Samples were prepared by spiking potential cross-reactive organisms into simulated nasopharyngeal swab sample matrix at the highest concentration possible based on the organism stock – at least 105 TCID50/ml for viral targets and 106 CFU/ml for bacterial and fungal targets. These concentrations represent levels approximately 800-1,000,000fold higher than the LoD of the QIAstat-Dx Respiratory Panel.

A certain level of cross-reactivity with off-panel Bordetella species and Bordetella pertussis was predicted by in silico sequence analysis and was observed when Bordetella holmesii and Bordetella bronchiseptica were tested in vitro.

24

Table 5.4: List of Analytical Specificity Pathogens

Pathogen Type	Pathogen
On-panel bacteria	Mycoplasma. pneumoniae
	Bordetella pertussis
	Chlamydia pneumoniae
Off-panel bacteria	Acinetobacter calcoaceticus
	Bordetella avium
	Bordetella bronchiseptica
	Bordetella hinzii
	Bordetella holmesii
	Bordetella parapertussis
	Chlamydia trachomatis
	Corynebacterium diphteriae
	Enterobacter aerogenes
	Escherichia coli (0157)
	Haemophilus influenzae
	Klebsiella oxytoca
	Klebsiella pneumoniae
	Lactobacillus acidophilus
	Lactobacillus plantarum
	Legionella bozemanii
	Legionella dumofii
	Legionella feeleii
	Legionella longbeacheae
	Legionella micdadei
	Legionella pneumophila
	Moraxella catarrhalis
	Mycobacterium tuberculosis*
	Mycoplasma genitalium
	Mycoplasma hominis
	Mycoplasma orale
	Neisseria elongata
	Neisseria gonorrhoeae
	Neisseria meningitidis
	Proteus mirabilis
	Pseudomonas aeruginosa
	Serratia marcescens
	Staphylococcus aureus
	Staphylococcus epidermidis
	Stenotrophomonas maltophilia
	Streptococcus agalactiae
	Streptococcus pneumoniae
	Streptococcus pyogenes
	Streptococcus salivarus
	Ureaplasma urealyticum

25

On-panel viruses	Influenza A H1N1
	Influenza A H3N2
	Influenza A H1N1/pdm09
	Influenza B
	Cor 229E
	Cor OC43
	Cor NL63
	Cor HKU1†
	Parainfluenza Virus 1
	Parainfluenza Virus 2
	Parainfluenza Virus 3
	Parainfluenza Virus 4a
	RSV A
	hMPV A
	Adenovirus C
	Adenovirus B
	Enterovirus
	Rhinovirus
Off-panel viruses	Bocavirus‡
	Cytomegalovirus
	Epstein-Barr Virus
	Herpes Simplex Virus 1
	Herpes Simplex Virus 2
	Measles Virus
	Middle East Respiratory Syndrome
	Coronavirus§
	Mumps
Off-panel fungi	Aspergillus flavus
	Aspergillus fumigatus
	Candida albicans
	Cryptococcus neoformans

Mycobacterium tuberculosis genomic DNA tested

† Coronavirus HKU1 clinical specimen tested

‡ Bocavirus Type 1 clinical specimens tested

§ Middle East Respiratory Syndrome Coronavirus synthetic RNA tested

Interference

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

26

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

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

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

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

None of the substances tested showed inhibition, except for the nasal influenza vaccines (Table 5.5). This was due to the fact that the selection of substances concentration was higher than the concentrations expected to be present in a sample. In addition, nasal influenza vaccines (Fluenz Tetra and FluMist) were predicted to be reactive with the QIAstat-Dx® Respiratory Panel Influenza A (subtype) and Influenza B assays. Final dilution without observable interfering effect was 0,00001% v/v for both vaccines.

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

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

Substance Tested	Concentration Tested	Results
Endogenous Substances
Human genomic DNA 200 ng/µL	20 ng/µL	No Interference
Human Blood (+NaCitrate)	1% v/v	No Interference
Mucin from bovine submaxillary	1% v/v	No Interference
Competitive Microorganisms
Staphylococcus aureus	1.00E+06 CFU/mL	No Interference
Neisseria meningitidis	5.00E+04 CFU/mL	No Interference
Corynebacterium diphtheriae	5.00E+03 CFU/mL	No Interference
Substance Tested	Concentration Tested	Results
Human Cytomegalovirus	1.00E+05 TCID50/mL	No Interference
Exogenous Substances
Tobramycin	0.6 mg/mL	No Interference
Mupirocin	2% w/v	No Interference
Saline Nasal Spray with Preservatives	1% v/v	No Interference
Afrin, Severe Congestion Nasal Spray
(Oxymetazoline HCl)	1% v/v	No Interference
Analgesic ointment
(Vicks®VapoRub®)	1% w/v	No Interference
Petroleum Jelly (Vaseline®)	1% w/v	No Interference
FluMist nasal influenza vaccine	0,00001% v/v	Interference
FluMist nasal influenza vaccine	0,000001% v/v	No Interference
Fluenz Tetra nasal influenza vaccine	0,00001% v/v	Interference
	0,000001% v/v	No Interference
Disinfecting/Cleaning Substances
Disinfecting wipes	½ inches²/1ml UTM	No Interference
DNAZap	1% v/v	No Interference
RNaseOUT	1% v/v	No Interference
Bleach	5% v/v	No Interference
Ethanol	5% v/v	No Interference
Specimen Collection Materials
Swab Copan 168C	1 swab/1mL UTM	No Interference
Swab Copan FloQ	1 swab/1mL UTM	No Interference
Swab Copan 175KS01	1 swab/1mL UTM	No Interference
Swab Puritan 25-801 A 50	1 swab/1mL UTM	No Interference
VTM Sigma Virocult	100%	No Interference
VTM Remel M4-RT	100%	No Interference
VTM Remel M4	100%	No Interference
VTM Remel M5	100%	No Interference
VTM Remel M6	100%	No Interference
BD Universal Viral Transport	100%	No Interference

Table 5.5: Final highest concentration without observable inhibitory effect.

27

28

Specimen Stability

Verification that storage of NPS samples at the specified conditions do not impact the performance when tested with the QIAstat-Dx® Respiratory Panel compared to freshly tested samples was evaluated. The detailed list of pathogens and strains for the 10 sample mixes used in the study is described in Table 5.6 with the respective 5x or 1xLoD concentration. Each pathogen was spiked into HeLa in UTM combined samples in a final concentration of 5x LoD or 1x LoD based on the 1x LoD concentration. During the study execution a total of 10 replicates per storage condition and target were tested.

| Mix | Pathogen | Strain | Source | Times
LoD | Final
Concentration
Stock titer
(re-titrated) |
|-----------|-------------------------|--------------------------------------|-------------|--------------|--------------------------------------------------------|
| Mix 1 | Influenza A H1 | A/New Caledonia/20/99 | Zeptometrix | 5x | 7.55E+6
TCID50/mL |
| | Cor HKU1* | n/a | Zeptometrix | 5x | n/a |
| | PIV2 | Greer | ATCC | 5x | 1.16E+7
TCID50/mL |
| | RSVB | CH93(18)-18 | Zeptometrix | 5x | 6.30E+06
TCID50/mL |
| | C. pneumoniae | TW183 | ATCC | 5x | 2.25E+06 IFU/mL |
| Mix 2 | Influenza B | B/Florida/4/2006 | ATCC | 5x | 5.40E+09
CEID50/mL |
| | Cor 229E | n/a | ATCC | 5x | 7.90E+04
TCID50/mL |
| | PIV4a** | M-25 | ATCC | 5x | 8.00E+04
TCID50/mL |
| | Enterovirus D68 | /US/IL/14-18952
(enterovirus D68) | ATCC | 5x | 4.45E+07
TCID50/mL |
| | hMPV A1 | hMPV-16, IA10-2003
(A1) | Zeptometrix | 5x | 7.55E+06
TCID50/mL |
| | B. pertussis | 1028 | ATCC | 5x | 1.35E+07 CFU/mL |
| Mix 3 | Influenza H1N1
(pdm) | A/Virginia/ATCC1/2009 | ATCC | 5x | 3.35E+06 PFU/mL |
| | Cor OC43 | n/a | ATCC | 5x | 1.41E+06
TCID50/mL |
| | PIV3 | C 243 | ATCC | 5x | 1.16E+07
TCID50/mL |
| | Rhinovirus A2 | HGP (rhinovirus A2) | ATCC | 5x | 1.41E+08
TCID50/mL |
| | RSVA | A2 | ATCC | 5x | 1.90E+08 PFU/mL |
| | M. pneumoniae | PI 1428 | ATCC | 5x | 5.00E+06 CU/mL |
| Mix | Pathogen | Strain | Source | Times
LoD | Final
Concentration
Stock titer
(re-titrated) |
| Mix 4 | Influenza A H3 | A/Port Chalmers/1/73 | ATCC | 5x | 7.90E+09
CEID50/mL |
| | Cor NL63*** | n/a | Zeptometrix | 5x | 5.85E+05
TCID50/mL |
| | PIV1 | C35 | ATCC | 5x | 2.50E+06
TCID50/mL |
| | Adenovirus B3 | GB (adenovirus B3) | ATCC | 5x | 7.90E+10
TCID50/mL |
| Mix 5 | Influenza A H1 | A/New Caledonia/20/99 | Zeptometrix | 1x | 1.51E+6
TCID50/mL |
| | Cor HKU1* | n/a | Zeptometrix | 1x | n/a |
| | PIV2 | Greer | ATCC | 1x | 2.32E+06
TCID50/mL |
| | RSVB | CH93(18)-18 | Zeptometrix | 1x | 1.26E+06
TCID50/mL |
| | C. pneumoniae | TW183 | ATCC | 1x | 4.50E+05 IFU/mL |
| Mix 6 | Influenza B | B/Florida/4/2006 | ATCC | 1x | 1.08E+09
CEID50/mL |
| | Cor 229E | n/a | ATCC | 1x | 1.58E+04
TCID50/mL |
| | PIV4a** | M-25 | ATCC | 1x | 1.60E+04
TCID50/mL |
| | Enterovirus D68 | /US/IL/14-18952
(enterovirus D68) | ATCC | 1x | 8.89E+06
TCID50/mL |
| | hMPV A1 | hMPV-16, IA10-2003
(A1) | Zeptometrix | 1x | 1.51E+06
TCID50/mL |
| Mix 7 | B. pertussis | 1028 | ATCC | 1x | 2.70E+06 CFU/mL |
| | Influenza H1N1
(pdm) | A/Virginia/ATCC1/2009 | ATCC | 1x | 6.70E+05 PFU/mL |
| | Cor OC43 | n/a | ATCC | 1x | 2.81E+05
TCID50/mL |
| | PIV3 | C 243 | ATCC | 1x | 2.32E+06
TCID50/mL |
| | Rhinovirus A2 | HGP (rhinovirus A2) | ATCC | 1x | 2.81E+07
TCID50/mL |
| | RSVA | A2 | ATCC | 1x | 3.80E+07 PFU/mL |
| | M. pneumoniae | PI 1428 | ATCC | 1x | 1.00E+06 CCU/mL |
| Mix | Pathogen | Strain | Source | Times
LoD | Final
Concentration
Stock titer
(re-titrated) |
| Mix 8 | Influenza A H3 | A/Port Chalmers/1/73 | ATCC | 1x | 1.58E+09
CEID50/mL |
| | Cor NL63*** | n/a | Zeptometrix | 1x | 1.17E+05
TCID50/mL |
| | PIV1 | C35 | ATCC | 1x | 5.00E+05
TCID50/mL |
| | Adenovirus B3 | GB (adenovirus B3) | ATCC | 1x | 1.58E+10
TCID50/mL |
| Mix 9 | Influenza A H1 | A/New Caledonia/20/99 | Zeptometrix | 1x | 1.51E+6
TCID50/mL |
| | Adenovirus B3 | GB (adenovirus B3) | ATCC | 1x | 1.58E+10
TCID50/mL |
| Mix
10 | Enterovirus D68 | /US/IL/14-18952
(enterovirus D68) | ATCC | 1x | 8.89E+06
TCID50/mL |
| | hMPV A1 | hMPV-16, IA10-2003
(A1) | Zeptometrix | 1x | 1.51E+06
TCID50/mL |
| | C. pneumoniae | TW183 | ATCC | 1x | 4.50E+05 IFU/mL |
| Mix 9 | Influenza A H1 | A/New Caledonia/20/99 | Zeptometrix | 1x | 1.51E+6
TCID50/mL |
| | Adenovirus B3 | GB (adenovirus B3) | ATCC | 1x | 1.58E+10
TCID50/mL |
| Mix
10 | Enterovirus D68 | /US/IL/14-18952
(enterovirus D68) | ATCC | 1x | 8.89E+06
TCID50/mL |
| | hMPV A1 | hMPV-16, IA10-2003
(A1) | Zeptometrix | 1x | 1.51E+06
TCID50/mL |
| | C. pneumoniae | TW183 | ATCC | 1x | 4.50E+05 IFU/mL |

Table 5.6: Pathogens tested in Specimen Stability Study

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The storage conditions are provided in Table 5.7.

Table 5.7: Storage conditions and samples tested per time point
------------------------------------------------------------------

| Storage
Condition | Time | Temperature | Samples tested with QIAstat-
Dx® Respiratory Panel |
|----------------------|---------|--------------|-------------------------------------------------------|
| Fresh | 0 h | 15 to 25 °C | Mix 1 to 8 |
| Condition 1 | 4 h | 15 to 25 °C | Mix 1 to 8 |
| Condition 2 | 72 h | 2-8 °C | Mix 1 to 8 |
| Condition 3 | 30 days | -15 to-25 °C | Mix 1 to 8 |

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Sample stability testing demonstrated that the QIAstat-Dx® Respiratory Panel Assay is capable of processing samples which are stored prior to the analysis under conditions typically utilized for NPS specimens according to the intended use.

The results of this study support the following recommendations for storage of NPS resuspended in UTM before testing:

Up to 4h at RT (15 to 25 °C).
Up to 3 days in the fridge (2 to 8 °C). .
Up to 30 days frozen (-15 to -25 °C). ●

Matrix Equivalency

A comparison of the performance of analytical samples prepared in NPS simulated matrix to negative clinical NPS sample matrix, and combined samples versus singlespiked samples was conducted. A total of 4 combined sample mixes were prepared by spiking individual pathogens in true-negative clinical NPS sample matrix for testing with QIAstat-Dx® Respiratory Panel. Every sample combination was established to detect not more than one positive pathogen per Reaction Chamber (RC). In order to assess comparable performance for the NPS clinical matrix, a concentration of 1x LoD for at least one strain per pathogen covering the QIAstat-Dx® Respiratory Panel was prepared in a true-negative clinical NPS sample matrix and tested in 20 replicates (using one or more lots of QIAstat-Dx® Respiratory Panel cartridges executed on one or more QIAstat-Dx® Analyzers). In addition, up to 6 pathogens were spiked per sample in order to demonstrate comparable performance to single-spiked samples (one analyte per sample). The LoD in clinical NPS sample matrix using combined samples was not shown to be equivalent to LoD in simulated matrix for all analytes (established with single-spiked samples). While claimed LoD concentrations represent the highest (most concentrated) titer of analyte confirmed in clinical matrix, analytical studies were performed in simulated matrix using the LoD determined in simulated matrix (the more challenging condition).

Reproducibility

Reproducibility testing of contrived samples was performed at three test sites. The study incorporated a range of potential variation factors introduced by sites, days, replicates, cartridge lots, operators, and QIAstat-Dx analyzers. For each site, testing was performed across 5 days with 4 replicates per day (leading to a total of 20 replicates per target, concentration and site), a minimum of 2 different QIAstat-Dx Analyzers per site, and at least 2 operators on each testing day.

A total of 12 sample mixes were prepared with at least 3 replicates tested per sample mix. Each pathogen was spiked into HeLa in UTM combined samples in a final concentration of 0.1x LoD, 1x LoD or 3x LoD, respectively. A summary of results for each analyte is provided in Table 5.8.

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Table 5.8 summarizes the results for 0.1x LoD concentration where it is observed that the detection rate for 24 of the 24 targets was