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.

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.

AI/ML Overview

The QIAGEN QIAstat-Dx Respiratory Panel is a multiplexed nucleic acid test intended 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). The device is intended for use with the QIAstat-Dx system to aid in the diagnosis of respiratory infections in individuals suspected of having such infections.

Acceptance Criteria and Device Performance

The acceptance criteria for the QIAstat-Dx Respiratory Panel are primarily demonstrated through its analytical performance (Limit of Detection, Analytical Reactivity, Analytical Specificity, Interference, Specimen Stability, and Reproducibility) and clinical performance (sensitivity and specificity compared to an FDA-cleared multiplexed respiratory pathogen panel). While explicit quantitative acceptance criteria for all aspects are not provided in the document (e.g., specific minimum acceptable sensitivity/specificity percentages), the data presented demonstrates the device meets acceptable performance levels for diagnostic assays.

Here's a table summarizing reported performance characteristics:

Performance Characteristic	Acceptance Criteria (Implied)	Reported Device Performance
Limit of Detection (LoD)	≥95% detection rate at specific concentrations for each pathogen	Achieved ≥95% (typically 19/20 or 20/20 positives) for all 51 pathogen strains at their determined LoD concentrations (e.g., Influenza A H1N1: 4 TCID50/ml, Coronavirus NL63: 0.01 TCID50/ml).
Analytical Reactivity (Inclusivity)	100% detection rate for clinically relevant and diverse strains at tested concentrations.	100% detection rate for all 127 tested influenza, coronavirus, parainfluenza, RSV, metapneumovirus, adenovirus, enterovirus, rhinovirus, Mycoplasma pneumoniae, Bordetella pertussis, and Chlamydophila pneumoniae strains at specified concentrations.
Analytical Specificity (Cross-Reactivity/Exclusivity)	No false positives due to common respiratory flora or other common pathogens, or clearly identified and disclosed cross-reactivity.	Only observed cross-reactivity with Bordetella holmesii and Bordetella bronchiseptica (off-panel Bordetella species), which was predicted by in silico analysis. No other significant cross-reactivity.
Interference	No inhibition or false results due to endogenous or exogenous substances (except for disclosed interferences like high concentrations of nasal influenza vaccines).	No inhibition observed except for high concentrations of nasal influenza vaccines (Fluenz Tetra and FluMist), which were predicted to be reactive and disclosed. Performance was unaffected by other substances.
Specimen Stability	Stable performance across specified storage conditions (RT, refrigerated, frozen).	Assay capable of processing samples stored up to 4 hours at RT, 3 days refrigerated, and 30 days frozen without impacting performance.
Reproducibility	Consistent detection rates across different sites, days, lots, operators, and analyzers. For concentrations near LoD (0.1x LoD), expected detection rate < 95%; for concentrations at or above LoD (1x LoD, 3x LoD), expected detection rate ≥ 95%.	0.1x LoD: All 24 targets showed <95% detection rates (e.g., Adenovirus: 50% overall, Bordetella pertussis: 42.37% overall). 1x LoD: All 24 targets showed ≥95% detection rates (e.g., Adenovirus: 100% overall, Bordetella pertussis: 96.67% overall). 3x LoD: All 24 targets showed ≥95% detection rates (e.g., Adenovirus: 100% overall, Bordetella pertussis: 100% overall).
Clinical Sensitivity (PPA)	High PPA against a comparator FDA-cleared multiplexed respiratory pathogen panel.	Overall PPA ranged from 0.0% (Influenza A H1, with only 1 positive sample in study) to 100.0% (Coronavirus HKU1, OC43, PIV1, PIV2, PIV4, Bordetella pertussis, Chlamydophila pneumoniae, Mycoplasma pneumoniae, though some with small denominators). Most targets demonstrated PPA > 85%, with many > 95%.
Clinical Specificity (NPA)	High NPA against a comparator FDA-cleared multiplexed respiratory pathogen panel.	Overall NPA consistently very high, ranging from 97.9% (Rhinovirus/Enterovirus) to 100.0% for all analytes.
Validity Rate	High rate of valid results on first attempt.	95.88% (1912/1994) valid results on first attempt in prospective study. 4.11% invalid/no result initially. After retesting, 72 of 82 initially failed specimens yielded valid results.

Study Details

This device is not an AI/ML powered device, therefore some of the questions (e.g. data provenance for test set, number of experts, adjudication method and MRMC study) are not relevant. This is a traditional IVD device and the study demonstrates the analytical and clinical performance through wet-lab and clinical sample testing against a comparator method.

A table of acceptance criteria and the reported device performance: See table above.
Sample sized used for the test set and the data provenance:
- Analytical Performance Studies (Non-clinical):
  - Limit of Detection: For each of the 51 pathogen strains, at least 20 replicates were tested. Data provenance is not explicitly stated as patient data, but rather contrived analytical samples prepared from commercial suppliers or artificial samples.
  - Analytical Reactivity: 127 respiratory pathogen isolates/strains were tested in triplicates.
  - Analytical Specificity: Samples were contrived by spiking off-panel organisms into simulated nasopharyngeal swab sample matrix.
  - Interference: 30 potentially interfering substances were tested in contrived samples (mix of organisms at 5xLoD and negative specimens).
  - Specimen Stability: 10 sample mixes (each containing multiple pathogens) were tested with 10 replicates per storage condition per target.
  - Matrix Equivalency: 20 replicates for at least one strain per pathogen, prepared in true-negative clinical NPS sample matrix.
  - Reproducibility: 12 sample mixes (contrived samples) were tested across 3 sites, with 20 replicates per target, concentration, and site (4 replicates/day for 5 days).
- Clinical Performance Study:
  - Sample Size: A total of 2,304 residual NPS specimens were tested: 1994 prospective (1093 frozen prospective, 901 fresh prospective) and 310 archived.
  - Data Provenance: The study was conducted at six (6) geographically diverse study sites, five (5) U.S. sites and one (1) international site. Specimens were collected from individuals suspected of respiratory tract infections. The study involved both prospective collection (patients meeting inclusion criteria) and archived specimens.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g. radiologist with 10 years of experience): Not applicable for this type of IVD device. The ground truth for the clinical study was established by an FDA-cleared multiplexed respiratory pathogen panel (the comparator method).
Adjudication method (e.g. 2+1, 3+1, none) for the test set: Not applicable as the ground truth was established by a single comparator device, not through human expert adjudication of images.
If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance: Not applicable. This is an IVD device, not an AI/ML-powered imaging device that assists human readers.
If a standalone (i.e. algorithm only without human-in-the-loop performance) was done: Yes, the device operates as a standalone diagnostic system, performing nucleic acid extraction, amplification, and detection without human interpretation beyond reading the displayed results. Its performance was directly compared to another FDA-cleared multiplexed respiratory pathogen panel.
The type of ground truth used (expert consensus, pathology, outcomes data, etc):
- Analytical Studies: Ground truth was established by known concentrations of pathogen strains (from commercial suppliers like ZeptoMetrix and ATCC) or artificial samples.
- Clinical Study: Ground truth was established by an FDA-cleared multiplexed respiratory pathogen panel, which served as the comparator method.
The sample size for the training set: Not applicable. This is a traditional IVD device, not an AI/ML product that undergoes a "training" phase with a large dataset. The device's design and analytical parameters are based on scientific principles of multiplex PCR and reagent formulation.
How the ground truth for the training set was established: Not applicable, as there is no "training set" in the context of an AI/ML model for this device.

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)

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 GmbHQIAGEN Strasse 1Hilden, Germany D-40724
Contact Person:	Melissa MahallSenior Director Regulatory Affairs19300 Germantown RoadGermantown, MD 20874
	Phone: 301-944-7768Email: 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

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.

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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

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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

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

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).

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.'sFilmArray® 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 Panelis a multiplexed nucleic acid testintended for use with QIAstat-Dxsystem for the simultaneousqualitative detection andidentification of multiplerespiratory viral and bacterialnucleic acids in nasopharyngealswabs (NPS) eluted in universaltransport media (UTM) obtainedfrom individuals suspected ofrespiratory tract infections. Thefollowing organism types andsubtypes are identified using theQIAstat-Dx Respiratory Panel:Adenovirus, Coronavirus 229E,Coronavirus HKU1, CoronavirusNL63, Coronavirus OC43, HumanMetapneumovirus A+B, InfluenzaA, Influenza A H1, Influenza AH3, Influenza A H1N1/pdm09,Influenza B, Parainfluenza Virus1, Parainfluenza Virus 2,Parainfluenza Virus 3,Parainfluenza Virus 4,Rhinovirus/Enterovirus,Respiratory Syncytial Virus A+B,Bordetella pertussis,Chlamydophila pneumoniae, andMycoplasma pneumoniae.	FilmArray® Respiratory Panel(RP) is a multiplexed nucleic acidtest intended for use with theFilmArray Instrument for thesimultaneous qualitative detectionand identification of multiplerespiratory viral and bacterialnucleic acids in nasopharyngealswabs (NPS) obtained fromindividuals suspected ofrespiratory tract infections. Thefollowing organism types andsubtypes are identified using theFilmArray RP: Adenovirus,Coronavirus 229E, CoronavirusHKU1, Coronavirus NL63,Coronavirus OC43, HumanMetapneumovirus, Influenza A,Influenza A subtype H1,Influenza A subtype H3,Influenza A subtype H1-2009,Influenza B, Parainfluenza Virus1, Parainfluenza Virus 2,Parainfluenza Virus 3,Parainfluenza Virus 4, HumanRhinovirus/Enterovirus,Respiratory Syncytial Virus,Bordetella pertussis,Chlamydophila pneumoniae, andMycoplasma pneumoniae. Thedetection and identification ofspecific viral and bacterial nucleicacids from individuals exhibitingsigns and symptoms of arespiratory infection aids in thediagnosis of respiratory infectionif used in conjunction with otherclinical and epidemiologicalinformation. The results of thistest should not be used as the solebasis for diagnosis, treatment, orother management decisions.
	The detection and identification ofspecific viral and bacterial nucleicacids from individuals presentingwith signs and symptoms of arespiratory infection aids in thediagnosis of respiratory infectionif used in conjunction with otherclinical and epidemiologicalinformation. The results of this testshould not be used as the solebasis for diagnosis, treatment, orother management decisions.	Negative results in the setting of a
Characteristic	Device	Predicate
	Negative results in the setting of arespiratory illness may be due toinfection with pathogens that arenot detected by the test or lowerrespiratory tract infection that isnot detected by a nasopharyngealswab specimen. Positive results donot rule out co-infection with otherorganisms: the agent(s) detectedby the QIAstat-Dx RespiratoryPanel may not be the definitecause of disease. Additionallaboratory testing (e.g. bacterialand viral culture,immunofluorescence, andradiography) may be necessarywhen evaluating a patient withpossible respiratory tract infection.	respiratory illness may be due toinfection with pathogens that arenot detected by this test or, lowerrespiratory tract infection that isnot detected by a nasopharyngealswab specimen. Positive resultsdo not rule out coinfection withother organisms: the agent(s)detected by the Film Array RPmay not be the definite cause ofdisease. Additional laboratorytesting (e.g. bacterial and viralculture, immunofluorescence, andradiography) may be necessarywhen evaluating a patient withpossible respiratory tractinfection.
	Due to the small number ofpositive specimens collected forcertain organisms during theprospective clinical study,performance characteristics forBordetella pertussis andParainfluenza Virus 1 wereestablished primarily withretrospective clinical specimens.	Due to the small number ofpositive specimens collected forcertain organisms during theprospective clinical study,performance characteristics forBordetella pertussis, Coronavirus229E, Coronavirus OC43,Influenza A H1, Influenza A H3,Influenza A H1-2009, InfluenzaB, Mycoplasma pneumoniae,Parainfluenza Virus 1,Parainfluenza Virus 2, andParainfluenza Virus 4 wereestablished primarily withretrospective clinical specimens.
	Performance characteristics forChlamydophila pneumoniae,Parainfluenza Virus 2,Parainfluenza Virus 4, Influenza Asubtype H1 and Coronavirus 229Ewere established primarily usingcontrived clinical specimens.	Performance characteristics forChlamydophila pneumoniae wereestablished primarily usingcontrived clinical specimens.
	Due to the genetic similaritybetween Human Rhinovirus andEnterovirus, the QIAstat-DxRespiratory Panel cannot reliablydifferentiate them. A positiveQIAstat-Dx Respiratory PanelRhinovirus/Enterovirus resultshould be followed up using an	Due to the genetic similaritybetween Human Rhinovirus andEnterovirus, the FilmArray RPcannot reliably differentiate them.A positive FilmArray RPRhinovirus/Enterovirus resultshould be followed-up using analternate method (e.g., cell culture
Characteristic	Device	Predicate
	alternate method (e.g., cell cultureor sequence analysis).Performance characteristics forInfluenza A were established whenInfluenza A H1N1-2009 and A H3were the predominant Influenza Aviruses in circulation. Performanceof detecting Influenza A may varyif other Influenza A strains arecirculating or a novel Influenza Avirus emerges. If infection with anovel Influenza A virus issuspected based on current clinicaland epidemiological screeningcriteria recommended by publichealth authorities, specimensshould be collected withappropriate infection controlprecautions for novel virulentInfluenza viruses and sent to stateor local health departments fortesting. Viral culture should not beattempted in these cases unless aBSL 3+ facility is available toreceive and culture specimens.	or sequence analysis).The FilmArray RP assay forCoronavirus OC43 may cross-react with some isolates ofCoronavirus HKU1. A dualpositive result may be due tocross-reactivity or may indicate aco-infection.Performance characteristics forInfluenza A were establishedwhen Influenza A H1-2009, AH1, and A H3 were thepredominant Influenza A virusesin circulation. Performance ofdetecting Influenza A may vary ifother Influenza A strains arecirculating or a novel Influenza Avirus emerges. If infection with anovel Influenza A virus issuspected based on currentclinical and epidemiologicalscreening criteria recommendedby public health authorities,specimens should be collectedwith appropriate infection controlprecautions for novel virulentInfluenza viruses and sent to stateor local health departments fortesting. Viral culture should notbe attempted in these cases unlessa BSL 3+ facility is available toreceive and culture specimens.
Specimen Type	Nasopharyngeal swabs (NPS)eluted in UTM	Nasopharyngeal swabs (NPS)
Assay TargetsAmplificationand DetectionTechnology	See analyte list above, RNA/ DNAPCR	See analyte list above, RNA/DNAPCR
Assay Controls	One internal control in eachcartridge to control for sampleprocessing that is subjected to allnucleic acid extraction and	Two controls are included in eachreagent pouch to control forsample processing and both stagesof PCR and melt analysis.
Characteristic	Device	Predicate
	amplification steps similar topatient samples. Labeling willrecommend use of negative andpositive external controlsregularly. Use transport mediumas the external Negative Control,and previously characterizedpositive samples or negativesample spiked with wellcharacterized target organisms asexternal Positive Controls.	Labeling recommends the use ofexternal positive and negativecontrols regularly. Use viraltransport medium as the externalnegative control, and previouslycharacterized positive samples ornegative samples spiked with wellcharacterized organisms asexternal positive controls.
	Differences
Nucleic AcidExtraction	Extraction of nucleic acids usingspin columns	Extraction of nucleic acids usingmagnetic beads
Amplificationand DetectionInstrumentSystem	QIAstat-Dx Analyzer	FilmArray Instrument

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

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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	Detectionrate
Influenza A H1N1	A/New Jersey/8/76	ATCC® VR-897	341 CEID50/ml	Flu A: 20/20H1: 20/20
	A/Brisbane/59/07	ZeptoMetrix®0810244CFHI	4 TCID50/ml	Flu A: 20/20H1: 20/20
	A/New Caledonia/20/99	ZeptoMetrix0810036CFHI	15 TCID50/ml	Flu A: 20/20H1: 19/20
Influenza A H3N2	A/Virginia/ATCC6/2012 *	ATCC VR-1811	0.1 PFU/ml	Flu A: 20/20H3: 20/20
	A/Wisconsin/67/2005 *	ZeptoMetrix0810252CFHI	3.8 TCID50/ml	Flu A: 20/20H3: 20/20
	A/Port Chalmers/1/73	ATCC VR-810	499.3CEID50/ml	Flu A: 20/20H3: 20/20
Influenza A,subtype H1N1/2009	A/Virginia/ATCC1/2009	ATCC VR-1736	67 PFU/ml	Flu A: 20/20H1N1: 20/20
	A/SwineNY/03/2009	ZeptoMetrix0810249CFHI	56 TCID50/ml	Flu A: 20/20H1N1: 20/20
Influenza B	B/Virginia/ATCC5/2012 *	ATCC VR-1807	0.03 PFU/ml	20/20
	B/FL/04/06	ATCC VR-1804	1080CEID50/ml	20/20
	B/Taiwan/2/62	ATCC VR-295	5000CEID50/ml	19/20
Coronavirus 229E	—	ATCC VR-740	0.2 TCID50/ml	20/20
	— *	ZeptoMetrix0810229CFHI	3.6 TCID50/ml	20/20
Coronavirus OC43	—	ATCC VR-1558	0.1 TCID50/ml	20/20
	— *	ZeptoMetrix0810024CFHI	0.1 TCID50/ml	20/20
Coronavirus NL63	—	ZeptoMetrix0810228CFHI	0.01TCID50/ml	20/20
Coronavirus HKU1	— *	Clinical SampleS510	40,000copies/ml	20/20
Parainfluenza Virus1 (PIV 1)	C35 *	ATCC VR-94	0.2 TCID50/ml	19/20
	—	ZeptoMetrix0810014CFHI	0.2 TCID50/ml	19/20
Pathogen	Strain	Source	Concentration	Detection rate
Parainfluenza Virus2 (PIV 2)	Greer	ATCC VR-92	7.3 TCID50/ml	20/20
Parainfluenza Virus2 (PIV 2)	- *	ZeptoMetrix0810015CFHI	1.3 TCID50/ml	19/20
Parainfluenza Virus3 (PIV 3)	C 243	ATCC VR-93	2.3 TCID50/ml	20/20
Parainfluenza Virus3 (PIV 3)	- *	ZeptoMetrix0810016CFHI	11.5 TCID50/ml	20/20
Parainfluenza Virus4a (PIV 4a)	M-25	ATCC VR-1378	0.5 TCID50/ml	20/20
Parainfluenza Virus4b (PIV 4b)	- *	ZeptoMetrix0810060BCFHI	9.5 TCID50/ml	20/20
RespiratorySyncytial Virus A	A2 *	ATCC VR-1540	12.0 PFU/ml	20/20
RespiratorySyncytial Virus A	Long *	ATCC VR-26	33.0 PFU/ml	20/20
RespiratorySyncytial Virus B	18537 *	ATCC VR-1580	0.03 PFU/ml	20/20
RespiratorySyncytial Virus B	CH93(18)-18	ZeptoMetrix0810040CFHI	0.4 TCID50/ml	20/20
HumanMetapneumovirus	Peru6-2003 (type B2) *	ZeptoMetrix0810159CFHI	0.01TCID50/ml	19/20
HumanMetapneumovirus	hMPV-16, IA10-2003(A1)	ZeptoMetrix0810161CFHI	0.5 TCID50/ml	20/20
HumanMetapneumovirus	hMPV-20, IA14-2003(A2) *	ZeptoMetrix,0810163CFHI	0.4 TCID50/ml	19/20
HumanMetapneumovirus	hMPV-3, Peru2-2002(B1) *	ZeptoMetrix,0810156CFHI	1479.9TCID50/ml	19/20
Adenovirus	GB (Adenovirus B3)	ATCC VR-3	4993.0TCID50/ml	20/20
Adenovirus	RI-67 (Adenovirus E4) *	ATCC VR-1572	15.8TCID50/ml	20/20
Adenovirus	Adenoid 75 (AdenovirusC5) *	ATCC VR-5	7331.0TCID50/ml	20/20
Adenovirus	Adenoid 71 (AdenovirusC1) *	ATCC VR-1	69.5TCID50/ml	20/20
Adenovirus	Adenoid 6 (AdenovirusC2) *	ATCC VR-846	28.1TCID50/ml	20/20
Adenovirus	Tonsil 99 (AdenovirusC6) *	ATCC VR-6	88.8TCID50/ml	20/20
Pathogen	Strain	Source	Concentration	Detectionrate
Enterovirus	/US/IL/14-18952(Enterovirus D68)	ATCC VR-1824	8.9TCID50/ml	19/20
	Echovirus 6 *	ATCC VR-241	0.9TCID50/ml	19/20
Rhinovirus	1059 (RhinovirusB14) *	ATCC VR-284	8.9TCID50/ml	20/20
	HGP (Rhinovirus A2)	ATCC VR-482	8.9TCID50/ml	19/20
	11757 (RhinovirusC16) *	ATCC VR-283	50.0TCID50/ml	20/20
	Type 1A *	ATCC VR-1559	8.9TCID50/ml	20/20
Mycoplasmapneumoniae	M129-B7 (type 1) *	ATCC 29342	0.1 CCU/ml	20/20
	PI 1428	ATCC 29085	1.0 CCU/ml	20/20
Chlamydiapneumoniae	TW183	ATCC VR-2282	14.2 IFU/ml	20/20
	CWL-029 *	ATCC VR-1310	120.0 IFU/ml	19/20
Bordetellapertussis	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

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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.

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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	CatalogueID	Concentrationtested	x-foldLoD
	MRC-2 (recombinantA/England/42/72 andA/PR/8/34 strains)	ATCC	VR-777	8891.4CEID50/mL	100xLoD
	A/Switzerland/9715293/2013	ATCC	VR-1837	1000CEID50/mL	1x LoD
	A/Virginia/ATCC1/2009a	ATCC	VR-1736	6.7 PFU/mL	1x LoD
	A/SwineNY/03/2009	Zeptometrix	0810249CFHI	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	100xLoD
Influenza AH1N1 pan	Swine NY/01/2009	Zeptometrix	0810248CFHI	138 TCID50/mL	0.3x LoD
	Swine NY/02/2009	Zeptometrix	0810109CFNHI	1.4 TCID50/mL	10x LoD
	A/California/07/2009NYMC X-179A	ATCC	VR-1884	1400CEID50/mL	0.1x LoD
	Canada/6294/09	Zeptometrix	0810109CFJHI	1.7 TCID50/mL	3x LoD
	Mexico/4108/09	Zeptometrix	0810166CFHI	14.1TCID50/mL	0.1x LoD
	Netherlands/2629/2009	BEIResources	NR-19823	16 TCID50/mL	0.3x LoD
Influenza AH2N2	Japan/305/1957 (nucleicacid)b	BEI	NR-2775	0.00326 RNAng/µL	1x LoD
	Korea/426/1968xPuertoRico/8/1934 (nucleicacid)b	BEI	NR-9679	0.0000625RNA ng/µL	0.3x LoD
Influenza AH5N3	A/Duck/Singapore/645/1997 (nucleic acid)b	BEI	NR-9682	0.002475 RNAng/µL	1x LoD
Influenza AH10N7	Chicken/Germany/N/49(nucleic acid)b	BEI	NR-2765	0.068 RNAng/µL	10x LoD
Pathogen	Strain	Supplier	CatalogueID	Concentrationtested	x-foldLoD
Influenza AH1N2	Recombinant KilbourneF63, A/NWS/1934 (HA)x A/RockefellerInstitute/5/1957 (nucleicacids) b	BEI	NR-9677	0.0148 RNAng/µL	100xLoD
	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	Notdetected
	B/Hong Kong/5/72 c	ATCC	VR-823	n/a	Notdetected
	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	2300CEID50/mL	3x LoD
	B/Florida/02/06 d	Zeptometrix	0810037CFHI	n/a	n/a
	B/Brisbane/60/2008	BEIResources	NR-42005	1.8 CEID50/mL	0.1x LoD
	B/Malaysia/2506/2004	BEIResources	NR-9723	1.58CEID50/mL	0.3x LoD
Coronavirus229E	n/a a	Zeptometrix	0810229CFHI	3.6 TCID50/mL	1x LoD
	n/a	ATCC	VR-740	0.2 TCID50/mL	0.3x LoD
CoronavirusOC43	n/a a	ATCC	VR-1558	0.1 TCID50/mL	1x LoD
	n/a	Zeptometrix	0810024CFHI	0.1 TCID50/mL	1x LoD
CoronavirusNL63	n/a a	Zeptometrix	0810228CFHI	0.01TCID50/mL	1x LoD
	n/a	BEIResources	NR-470	1.6 TCID50/mL	1x LoD
Pathogen	Strain	Supplier	CatalogueID	Concentrationtested	x-foldLoD
CoronavirusHKU1	n/a a, e	Zeptometrix	NATRVP-IDI	3E+03copies/mL	1x LoD
	n/a e	QIAGENBarcelona(STAT-Dx)	ClinicalsampleS510	1.2E+04copies/mL	0.3x LoD
	n/a e	QIAGENBarcelona(STAT-Dx)	ClinicalsampleS501	7E+03copies/mL	1x LoD
	n/a e	QIAGENBarcelona(STAT-Dx)	ClinicalsampleS496	7E+03copies/mL	1x LoD
ParainfluenzaVirus 1	n/a a	Zeptometrix	0810014CFHI	0.02TCID50/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
Parainfluenza Virus 2	n/a	Zeptometrix	0810015CFHI	1.3 TCID50/mL	0.3x LoD
	n/a	Zeptometrix	0810504CFHI	1.3 TCID50/mL	0.1x LoD
Parainfluenza Virus 3	n/a a	Zeptometrix	0810016CFHI	11.5TCID50/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
Parainfluenza Virus 4	M-25 a	ATCC	VR-1378	0.5 TCID50/mL	1x LoD
	n/a	Zeptometrix	0810060BCFHI	9.6 TCID50/mL	0.3x LoD
	n/a	Zeptometrix	0810060CFHI	28.2TCID50/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
RespiratorySyncytialVirus A+B	CH93(18)-18	Zeptometrix	0810040CFHI	0.4 TCID50/mL	1x LoD
	n/a	Zeptometrix	0810040ACFHI	0.3 TCID50/mL	0.1x LoD
	B WV/14617/85	ATCC	VR-1400	15.8TCID50/mL	1x LoD
HumanMetapneumovirus	IA10-2003 a	Zeptometrix	0810161CFHI	0.5 TCID50/mL	1x LoD
	IA14-2003	Zeptometrix	0810163CFHI	0.4 TCID50/mL	1x LoD
Pathogen	Strain	Supplier	CatalogueID	Concentrationtested	x-foldLoD
	Peru2-2002	Zeptometrix	0810156CFHI	1478.9TCID50/mL	1x LoD
	Peru6-2003	Zeptometrix	0810159CFHI	0.01TCID50/mL	1x LoD
	IA3-2002	Zeptometrix	0810160CFHI	66 TCID50/mL	3x LoD
	IA27-2004	Zeptometrix	0810164CFHI	1.3 TCID50/mL	1x LoD
	Peru3-2003	Zeptometrix	0810158CFHI	31.6TCID50/mL	1x LoD
	IA18-2003	Zeptometrix	0810162CFHI	0.4 TCID50/mL	1x LoD
	Peru1-2002	Zeptometrix	0810157CFHI	2187.8TCID50/mL	10x LoD
Adenovirus	Tonsil 99 a	ATCC	VR-6	88.8TCID50/mL	1x LoD
	GB	ATCC	VR-3	4992.8TCID50/mL	0.3x LoD
	Adenoid 71	ATCC	VR-1	69.5TCID50/mL	1x LoD
	Adenoid 6	ATCC	VR-846	28.1TCID50/mL	0.3x LoD
	Adenoid 75	ATCC	VR-5	7331.2TCID50/mL	0.3x LoD
	RI-67	ATCC	VR-1572	15.8TCID50/mL	0.3x LoD
	Huie	ATCC	VR-863	88.9TCID50/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	0.28TCID50/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	28117TCID50/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	889.1TCID50/mL	10x LoD
Pathogen	Strain	Supplier	CatalogueID	Concentrationtested	x-foldLoD
	Bastianni	ATCC	VR-1660	281.2TCID50/mL	1x LoD
	Griggs	ATCC	VR-1311	1.6 TCID50/mL	0.3x LoD
	Conn-5	ATCC	VR-28	158.1TCID50/mL	0.3x LoD
	Ohio-1	ATCC	VR-29	2811.7TCID50/mL	3x LoD
	Nancy	ATCC	VR-30	0.9 TCID50/mL	0.3x LoD
	CHHE-29	ATCC	VR-47	0.03TCID50/mL	10x LoD
	Kuykendall [V-024-001-012]	ATCC	VR-850	28.1TCID50/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	49.9TCID50/mL	0.3x LoD
	FEB	ATCC	VR-483	281.2TCID50/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.

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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.

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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	TimesLoD	FinalConcentrationStock titer(re-titrated)
Mix 1	Influenza A H1	A/New Caledonia/20/99	Zeptometrix	5x	7.55E+6TCID50/mL
	Cor HKU1*	n/a	Zeptometrix	5x	n/a
	PIV2	Greer	ATCC	5x	1.16E+7TCID50/mL
	RSVB	CH93(18)-18	Zeptometrix	5x	6.30E+06TCID50/mL
	C. pneumoniae	TW183	ATCC	5x	2.25E+06 IFU/mL
Mix 2	Influenza B	B/Florida/4/2006	ATCC	5x	5.40E+09CEID50/mL
	Cor 229E	n/a	ATCC	5x	7.90E+04TCID50/mL
	PIV4a**	M-25	ATCC	5x	8.00E+04TCID50/mL
	Enterovirus D68	/US/IL/14-18952(enterovirus D68)	ATCC	5x	4.45E+07TCID50/mL
	hMPV A1	hMPV-16, IA10-2003(A1)	Zeptometrix	5x	7.55E+06TCID50/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+06TCID50/mL
	PIV3	C 243	ATCC	5x	1.16E+07TCID50/mL
	Rhinovirus A2	HGP (rhinovirus A2)	ATCC	5x	1.41E+08TCID50/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	TimesLoD	FinalConcentrationStock titer(re-titrated)
Mix 4	Influenza A H3	A/Port Chalmers/1/73	ATCC	5x	7.90E+09CEID50/mL
	Cor NL63***	n/a	Zeptometrix	5x	5.85E+05TCID50/mL
	PIV1	C35	ATCC	5x	2.50E+06TCID50/mL
	Adenovirus B3	GB (adenovirus B3)	ATCC	5x	7.90E+10TCID50/mL
Mix 5	Influenza A H1	A/New Caledonia/20/99	Zeptometrix	1x	1.51E+6TCID50/mL
	Cor HKU1*	n/a	Zeptometrix	1x	n/a
	PIV2	Greer	ATCC	1x	2.32E+06TCID50/mL
	RSVB	CH93(18)-18	Zeptometrix	1x	1.26E+06TCID50/mL
	C. pneumoniae	TW183	ATCC	1x	4.50E+05 IFU/mL
Mix 6	Influenza B	B/Florida/4/2006	ATCC	1x	1.08E+09CEID50/mL
	Cor 229E	n/a	ATCC	1x	1.58E+04TCID50/mL
	PIV4a**	M-25	ATCC	1x	1.60E+04TCID50/mL
	Enterovirus D68	/US/IL/14-18952(enterovirus D68)	ATCC	1x	8.89E+06TCID50/mL
	hMPV A1	hMPV-16, IA10-2003(A1)	Zeptometrix	1x	1.51E+06TCID50/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+05TCID50/mL
	PIV3	C 243	ATCC	1x	2.32E+06TCID50/mL
	Rhinovirus A2	HGP (rhinovirus A2)	ATCC	1x	2.81E+07TCID50/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	TimesLoD	FinalConcentrationStock titer(re-titrated)
Mix 8	Influenza A H3	A/Port Chalmers/1/73	ATCC	1x	1.58E+09CEID50/mL
	Cor NL63***	n/a	Zeptometrix	1x	1.17E+05TCID50/mL
	PIV1	C35	ATCC	1x	5.00E+05TCID50/mL
	Adenovirus B3	GB (adenovirus B3)	ATCC	1x	1.58E+10TCID50/mL
Mix 9	Influenza A H1	A/New Caledonia/20/99	Zeptometrix	1x	1.51E+6TCID50/mL
	Adenovirus B3	GB (adenovirus B3)	ATCC	1x	1.58E+10TCID50/mL
Mix10	Enterovirus D68	/US/IL/14-18952(enterovirus D68)	ATCC	1x	8.89E+06TCID50/mL
	hMPV A1	hMPV-16, IA10-2003(A1)	Zeptometrix	1x	1.51E+06TCID50/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+6TCID50/mL
	Adenovirus B3	GB (adenovirus B3)	ATCC	1x	1.58E+10TCID50/mL
Mix10	Enterovirus D68	/US/IL/14-18952(enterovirus D68)	ATCC	1x	8.89E+06TCID50/mL
	hMPV A1	hMPV-16, IA10-2003(A1)	Zeptometrix	1x	1.51E+06TCID50/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
------------------------------------------------------------------

StorageCondition	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

{31}------------------------------------------------

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.

{32}------------------------------------------------

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 <95% and therefore the acceptance criteria is met.

Target(0.1xLoD)	Site	Detection Rate(#Positive)	% Detection rate(#Positive)	95% Confidence Interval
Adenovirus(ATCC VR-3)	STAT	10 / 20	50.00%	29.9-70.1%
	LACNY	9 / 19	47.37%	27.3-68.2%
	INDIANA	10 / 19	52.63%	31.7-72.7%
	All Sites(Overall)	29 / 58	50.00%	37.5-62.5%
B. pertussis(BAA-2707)	STAT	9 / 20	45.00%	25.8-65.8%
	LACNY	7 / 19	36.84%	19.2-59.0%
	INDIANA	9 / 20	45.00%	25.8-65.8%
	All Sites(Overall)	25 / 59	42.37%	30.6-55.1%
C. pneumoniae(ATCC VR-2282)	STAT	11 / 20	55.00%	34.2-74.2%
	LACNY	11 / 19	57.89%	36.3-76.9%
	INDIANA	14 / 20	70.00%	48.1-85.5%
	All Sites(Overall)	36 / 59	61.02%	48.3-72.4%
Coronavirus229E (ATCC VR-740)	STAT	9 / 20	45.00%	25.8-65.8%
	LACNY	12 / 19	63.16%	41.0-80.9%
	INDIANA	5 / 20	25.00%	11.2-46.9%
	All Sites(Overall)	26 / 59	44.07%	32.2-56.7%
CoronavirusHKU1(NATRVP-IDI)	STAT	17 / 20	85.00%	64.0-94.8%
	LACNY	10 / 19	52.63%	31.7-72.7%
	INDIANA	9 / 20	45.00%	25.8-65.8%
	All Sites(Overall)	36 / 59	61.02%	48.3-72.4%
CoronavirusNL63(0810228CFHI)	STAT	13 / 20	65.00%	43.3-81.9%
	LACNY	12 / 19	63.16%	41.0-80.9%
	INDIANA	14 / 19	73.68%	51.2-88.2%
	All Sites(Overall)	39 / 58	67.24%	54.4-77.9%

Target	Site	%	95% Confidence Interval
Target 1	Site 1	100	100-100
	Site 2	100	100-100
	Site 3	100	100-100
Target 2	Site 1	100	100-100
	Site 2	100	100-100
	Site 3	100	100-100
Target 3	Site 1	100	100-100
	Site 2	100	100-100
	Site 3	100	100-100

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Page 30 of 43

Target(0.1xLoD)	Site	DetectionRate(#Positive)	% Detectionrate(#Positive)	95%ConfidenceInterval
CoronavirusOC43(ATCC VR-1558)	STAT	13 / 20	65.00%	43.3-81.9%
	LACNY	15 / 20	75.00%	53.1-88.8%
	INDIANA	15 / 20	75.00%	53.1-88.8%
	All Sites(Overall)	43 / 60	71.67%	59.2-81.5%
Enterovirus(ATCC VR-1824)	STAT	8 / 20	40.00%	21.9-61.3%
	LACNY	6 / 19	31.58%	15.4-54.0%
	INDIANA	7 / 20	35.00%	18.1-56.7%
	All Sites(Overall)	21 / 59	35.59%	24.6-48.3%
HumanMetapneumovirus (0810161CF)	STAT	6 / 20	30.00%	14.6-51.9%
	LACNY	9 / 19	47.37%	27.3-68.2%
	INDIANA	9 / 20	45.00%	25.8-65.8%
	All Sites(Overall)	24 / 59	40.68%	29.1-53.4%
Influenza A(0810249CFHI)	STAT	19 / 20	95.00%	76.4-99.1%
	LACNY	18 / 20	90.00%	69.9-97.2%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	57 / 60	95.00%	86.3-98.3%
Influenza A(ATCC VR-810)	STAT	10 / 20	50.00%	29.9-70.1%
	LACNY	9 / 19	47.37%	27.3-68.3%
	INDIANA	16 / 19	84.21%	62.4-94.5%
	All Sites(Overall)	35 / 58	60.34%	47.5-71.9%
Influenza A(ATCC VR-897)	STAT	14 / 20	70.00%	48.1-85.5%
	LACNY	9 / 19	47.37%	27.3-68.3%
	INDIANA	12 / 20	60.00%	38.7-78.1%
	All Sites(Overall)	35 / 59	59.32%	46.6-70.9%
Influenza AH1(ATCC VR-897)	STAT	13 / 20	65.00%	43.3-81.9%
	LACNY	13 / 19	68.42%	46.0-84.6%
	INDIANA	15 / 20	75.00%	53.1-88.8%
	All Sites(Overall)	41 / 59	69.49%	56.9-79.8%
Target(0.1xLoD)	Site	DetectionRate(#Positive)	% Detectionrate(#Positive)	95%ConfidenceInterval
Influenza B(ATCC VR-295)	STAT	7 / 20	35.00%	18.1-56.7%
	LACNY	9 / 19	47.37%	27.3-68.3%
	INDIANA	8 / 20	40.00%	21.9-61.3%
	All Sites(Overall)	24 / 59	40.68%	29.1-53.4%
Influenza H1N1(pdm09)(0810249CFHI)	STAT	14 / 20	70.00%	48.1-85.5%
	LACNY	16 / 20	80.00%	58.4-91.9%
	INDIANA	15 / 20	75.00%	53.1-88.8%
	All Sites(Overall)	45 / 60	75.00%	62.8-84.2%
Influenza H3(ATCC VR-810)	STAT	13 / 20	65.00%	43.3-81.9%
	LACNY	16 / 19	84.21%	62.4-94.5%
	INDIANA	17 / 19	89.47%	68.6-97.1%
	All Sites(Overall)	46 / 58	79.31%	67.2-87.8%
Mycoplasmapneumoniae(29085)	STAT	13 / 20	65.00%	43.3-81.9%
	LACNY	14 / 20	70.00%	48.1-85.5%
	INDIANA	14 / 20	70.00%	48.1-85.5%
	All Sites(Overall)	41 / 60	68.33%	55.8-78.7%
Parainfluenzavirus 1(0810014CFHI)	STAT	14 / 20	70.00%	48.1-85.5%
	LACNY	12 / 19	63.16%	41.0-80.9%
	INDIANA	9 / 19	47.37%	27.3-68.3%
	All Sites(Overall)	35 / 58	60.34%	47.5-71.9%
Parainfluenzavirus 2 (ATCCVR-92)	STAT	9 / 20	45.00%	25.8-65.8%
	LACNY	11 / 19	57.89%	36.3-76.9%
	INDIANA	12 / 20	60.00%	38.7-78.1%
	All Sites(Overall)	32 / 59	54.24%	41.7-66.3%
Parainfluenzavirus 3 (ATCCVR-93)	STAT	13 / 20	65.00%	43.3-81.9%
	LACNY	17 / 20	85.00%	64.0-94.8%
	INDIANA	17 / 20	85.00%	64.0-94.8%
	All Sites(Overall)	47 / 60	78.33%	66.4-86.9%
Target(0.1xLoD)	Site	DetectionRate(#Positive)	%Detectionrate(#Positive)	95%ConfidenceInterval
Parainfluenzavirus 4 (ATCCVR-1378)	STAT	10 / 20	50.00%	29.9-70.1%
	LACNY	11 / 19	57.89%	36.3-76.9%
	INDIANA	9 / 20	45.00%	25.8-65.8%
	All Sites(Overall)	30 / 59	50.85%	38.4-63.2%
RSVA (ATCCVR-1540)	STAT	6 / 20	30.00%	14.6-51.9%
	LACNY	7 / 20	35.00%	18.1-56.7%
	INDIANA	9 / 20	45.00%	25.8-65.8%
	All Sites(Overall)	22 / 60	36.67%	25.6-49.3%
RespiratorySyncytial VirusB (0810040CF)	STAT	14 / 20	70.00%	48.1-85.5%
	LACNY	15 / 19	78.95%	56.7-91.5%
	INDIANA	10 / 20	50.00%	29.9-70.1%
	All Sites(Overall)	39 / 59	66.10%	53.4-76.9%
Rhinovirus(ATCC VR-482)	STAT	15 / 20	75.00%	53.1-88.8%
	LACNY	15 / 20	75.00%	53.1-88.8%
	INDIANA	18 / 20	90.00%	69.9-97.2%
	All Sites(Overall)	48 / 60	80.00%	68.2-88.2%

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Table 5.9 summarizes the results for 1x LoD concentration where it is observed that the detection rate for 24 of the 24 targets was ≥95% and therefore the acceptance criteria is met.

		Table 5.9: Detection rate per target at 1x LoD concentration for each site of
		reproducibility study and 2-sided 95% Confidence Interval by target

Target (1xLoD)	Site	DetectionRate(#Positive)	0/0Detectionrate(#Positive)	તે જેને જીરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં લોકોનો મુખ્ય વ્યવસાય ખેતી, ખેતમજૂરી તેમ જ પશુપાલન છે. આ ગામમાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલConfidenceInterval
Adenovirus	STAT	20 / 20	100.00%	83.9-100%
(ATCC VR-3)	LACNY	18 / 18	100.00%	82.4-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	૨૪ ૨૪	100.00%	93.8-100%
Target (1xLoD)	Site	Detection Rate (#Positive)	% Detection rate (#Positive)	95% Confidence Interval
B. pertussis (ATCC BAA-2707)	STAT	18 / 20	90.00%	69.9-97.2%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	58 / 60	96.67%	88.6-99.1%
C. pneumoniae (ATCC VR-2282)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	60 / 60	100.00%	94.0-100%
Coronavirus 229E (ATCC VR-740)	STAT	18 / 20	90.00%	69.9-97.2%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	58 / 60	96.67%	88.6-99.1%
Coronavirus HKU1 (NATRVP-IDI)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	60 / 60	100.00%	94.0-100%
Coronavirus NL63 (0810228CFHI)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	18 / 18	100.00%	82.4-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	58 / 58	100.00%	93.8-100%
Coronavirus OC43 (ATCC VR-1558)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	59 / 59	100.00%	93.9-100%
Enterovirus (ATCC VR-1824)	STAT	19 / 20	95.00%	76.4-99.1%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	19 / 20	95.00%	76.4-99.1%
	All Sites (Overall)	58 / 60	96.67%	88.6-99.1%
Human Metapneumovirus (0810161CF)	STAT	19 / 20	95.00%	76.4-99.1%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	59 / 60	98.33%	91.1-99.7%
Target (1xLoD)	Site	Detection Rate(#Positive)	% Detection rate(#Positive)	95% Confidence Interval
Influenza A(0810249CFHI)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%
Influenza A(ATCC VR-810)	STAT	19 / 20	95.00%	76.4-99.1%
	LACNY	18 / 18	100.00%	82.4-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	57 / 58	98.28%	90.9-99.7%
Influenza A(ATCC VR-897)	STAT	19 / 20	95.00%	76.4-99.1%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 60	98.33%	91.1-99.7%
Influenza AH1(ATCC VR-897)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	19 / 20	95.00%	76.4-99.1%
	All Sites(Overall)	59 / 60	98.33%	91.1-99.7%
Influenza B(ATCC VR-295)	STAT	19 / 20	95.00%	76.4-99.1%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 60	98.33%	91.1-99.7%
Influenza H1N1(pdm09)(0810249CFHI)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%
Influenza H3(ATCC VR-810)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	18 / 18	100.00%	82.4-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	58 / 58	100.00%	93.8-100%
Mycoplasmapneumoniae(ATCC 29085)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%
Target (1xLoD)	Site	Detection Rate(#Positive)	% Detection rate(#Positive)	95% Confidence Interval
Parainfluenzavirus 1(0810014CFHI)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	18 / 18	100.00%	82.4-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	58 / 58	100.00%	93.8-100%
Parainfluenzavirus 2 (ATCCVR-92)	STAT	19 / 20	95.00%	76.4-99.1%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	19 / 20	95.00%	76.4-99.1%
	All Sites(Overall)	58 / 60	96.67%	88.6-99.1%
Parainfluenzavirus 3 (ATCCVR-93)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%
Parainfluenzavirus 4 (ATCCVR-1378)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	60 / 60	100.00%	94.0-100%
RSVA (ATCCVR-1540)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%
RespiratorySyncytial VirusB (0810040CF)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	60 / 60	100.00%	94.0-100%
Rhinovirus(ATCC VR-482)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%

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Table 5.10 summarizes the results for 3x LoD concentration where it is observed that the detection rate for 24 of the 24 targets was ≥95% and therefore the acceptance criteria has been met.

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Table 5.10: Detection rate per target at 3x LoD concentration for each site of
reproducibility study and 2-sided 95% Confidence Interval by target.

Target	Site	Detection Rate (#Positive)	% Detection rate (#Positive)	95% Confidence Interval
Adenovirus(ATCC VR-3)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	59 / 59	100.00%	93.9-100%
B. pertussis(ATCC BAA-2707)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	59 / 59	100.00%	93.9-100%
C. pneumoniae(ATCC VR-2282)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 20	95.00%	76.4-99.1%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	59 / 60	98.33%	91.1-99.7%
Coronavirus 229E (ATCC VR-740)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	59 / 59	100.00%	93.9-100%
Coronavirus HKU1 (NATRVP-IDI)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	60 / 60	100.00%	94.0-100%
Coronavirus NL63 (0810228CFHI)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites (Overall)	59 / 59	100.00%	93.9-100%
Target	Site	Detection Rate(#Positive)	% Detection rate(#Positive)	95% Confidence Interval
CoronavirusOC43 (ATCCVR-1558)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	19 / 19	100.00%	83.2-100%
	All Sites(Overall)	58 / 58	100.00%	93.8-100%
Enterovirus(ATCC VR-1824)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%
HumanMetapneumovirus (0810161CF)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%
Influenza A(0810249CFHI)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	19 / 19	100.00%	83.2-100%
	All Sites(Overall)	58 / 58	100.00%	93.8-100%
Influenza A(ATCC VR-810)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%
Influenza A(ATCC VR-897)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	60 / 60	100.00%	94.0-100%
Influenza AH1(ATCC VR-897)	STAT	19 / 20	95.00%	76.4-99.1%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 60	98.33%	91.1-99.7%
Influenza B(ATCC VR-295)	STAT	19 / 20	95.00%	76.4-99.1%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	58 / 59	98.31%	91.0-99.7%
Target	Site	Detection Rate(#Positive)	% Detection rate(#Positive)	95% Confidence Interval
Influenza H1N1(pdm09)(0810249CFHI)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	19 / 19	100.00%	83.2-100%
	All Sites(Overall)	58 / 58	100.00%	93.8-100%
Influenza H3(ATCC VR-810)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%
Mycoplasmapneumoniae(ATCC 29085)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	19 / 19	100.00%	83.2-100%
	All Sites(Overall)	58 / 58	100.00%	93.8-100%
Parainfluenzavirus 1(0810014CFHI)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%
Parainfluenzavirus 2 (ATCCVR-92)	STAT	19 / 20	95.00%	76.4-99.1%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 60	98.33%	91.1-99.7%
Parainfluenzavirus 3 (ATCCVR-93)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	19 / 19	100.00%	83.2-100%
	All Sites(Overall)	58 / 58	100.00%	93.8-100%
Parainfluenzavirus 4 (ATCCVR-1378)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	59 / 59	100.00%	93.9-100%
Target	Site	DetectionRate(#Positive)	%Detectionrate(#Positive)	95%ConfidenceInterval
RSVA (ATCCVR-1540)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	19 / 19	100.00%	83.2-100%
	All Sites(Overall)	58 / 58	100.00%	93.8-100%
RespiratorySyncytial VirusB (0810040CF)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	20 / 20	100.00%	83.9-100%
	INDIANA	20 / 20	100.00%	83.9-100%
	All Sites(Overall)	60 / 60	100.00%	94.0-100%
Rhinovirus(ATCC VR-482)	STAT	20 / 20	100.00%	83.9-100%
	LACNY	19 / 19	100.00%	83.2-100%
	INDIANA	19 / 19	100.00%	83.2-100%
	All Sites(Overall)	58 / 58	100.00%	93.8-100%

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Performance Characteristics - Clinical Studies

The clinical performance of the QIAstat-Dx Respiratory Panel was established during a multi-center study conducted at six (6) geographically diverse study sites: five (5) U.S. sites and one (1) international site. Each study location was representative of the intended use setting (clinical laboratories) and testing was performed by trained clinical laboratory personnel. Residual nasopharyngeal swab (NPS) samples were collected from subjects with signs and symptoms of respiratory infection for QIAstat-Dx Respiratory Panel and comparator testing.

A residual NPS specimens in UTM from each study subject was tested with the OLAstat-Dx Respiratory Panel and the comparator FDA cleared multiplexed respiratory pathogen panel, that matched all panel members, in accordance with product instructions for use. Specimens tested in the clinical study were collected using the Universal Transport Medium (UTM™) (Copan Diagnostics (Brescia, Italy and CA, USA)), MicroTest™ 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) and UniTranz-RT® Universal Transport Media (Puritan Diagnostics, ME , USA) collection kits.

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

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(N=1,093). No frozen samples were distributed amongst sites. At time of testing, specimens were thawed and tested on both the OIAstat-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. The performance of the OIAstat-Dx Respiratory Panel was evaluated by comparing the QIAstat-Dx Respiratory Panel test results with those from an FDA-cleared multiplexed respiratory pathogen panel.

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

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

The QIAstat-Dx Respiratory Panel prospective performance data in positive percent and negative percent agreements against the comparator methods are presented by analyte in Table 5.11.

Analyte		TP/(TP +FN)	Sensitivity /PPA	95% CI	TN/(TN+FP)	Specificity /NPA	95% CI
Viruses
Adenovirusa	Fresh	55/58	94.8%	85.9 - 98.2	833/839	99.3%	98.4 - 99.7
	Frozen	31/32	96.9%	84.3 - 99.4	1047/1057	99.1%	98.3 - 99.5
	Overall	86/90	95.6%	89.1 - 98.3	1880/1896	99.2%	98.6 - 99.5
Analyte		TP/(TP+FN)	Sensitivity/PPA	95% CI	TN/(TN+FP)	Specificity/NPA	95% CI
Coronavirus 229E	Fresh	8/9	88.9%	56.5-98.0	886/886	100.0%	99.6-100.0
	Frozen	0/0	N/A	N/A	1089/1089	100.0%	99.6-100.0
	Overall	8/9	88.9%	56.5-98.0	1975/1975	100.0%	99.8-100.0
Coronavirus HKU1b	Fresh	3/3	100.0%	43.8-100.0	890/892	99.8%	99.2-99.9
	Frozen	48/49	98.0%	89.3-99.6	1035/1040	99.5%	98.9-99.8
	Overall	51/52	98.1%	89.9-99.7	1925/1932	99.6%	99.3-99.8
Coronavirus NL63c	Fresh	4/5	80.0%	37.6-96.4	890/890	100.0%	99.6-100.0
	Frozen	36/42	85.7%	72.2-93.3	1046/1048	99.8%	99.3-99.9
	Overall	40/47	85.1%	72.3-92.6	1936/1938	99.9%	99.6-100.0
Coronavirus OC43d	Fresh	3/3	100.0%	43.8-100.0	892/892	100.0%	99.6-100.0
	Frozen	23/26	88.5%	71.0-96.0	1059/1063	99.6%	99.0-99.9
	Overall	26/29	89.7%	73.6-96.4	1951/1955	99.8%	99.5-99.9
Human Metapneumoviruse	Fresh	62/67	92.5%	83.7-96.8	828/829	99.9%	99.3-100.0
	Frozen	53/55	96.4%	87.7-99.0	1030/1034	99.6%	99.0-99.8
	Overall	115/122	94.3%	88.6-97.2	1858/1863	99.7%	99.4-99.9
Rhinovirus/Enterovirusf	Fresh	144/157	91.7%	86.3-95.1	715/739	96.7%	95.2-97.8
	Frozen	124/137	90.5%	84.4-94.4	941/953	98.7%	97.8-99.3
	Overall	268/294	91.2%	87.4-93.9	1656/1692	97.9%	97.1-98.5
Influenza Ag	Fresh	132/133	99.2%	95.8-99.9	753/757	99.5%	98.6-99.8
	Frozen	110/111	99.1%	95.1-99.8	972/977	99.5%	98.8-99.8
	Overall	242/244	99.2%	97.0-99.8	1725/1734	99.5%	99.0-99.7
Influenza A H1h	Fresh	0/1	0.0%	0.0-79.3	894/894	100.0%	99.6-100.0
	Frozen	0/0	N/A	N/A	1089/1089	100.0%	99.6-100.0
	Overall	0/1	0.0%	0.0-79.3	1983/1983	100.0%	99.8-100.0
Influenza A H1N1/pdm09i	Fresh	62/63	98.4%	91.5-99.78	826/831	99.4%	98.6-99.7
	Frozen	18/18	100.0%	82.4-100.0	1071/1071	100.0%	99.6-100.0
	Overall	80/81	98.8%	93.3-99.8	1897/1902	99.7%	99.4-99.9
Influenza A H3j	Fresh	67/67	100.0%	94.5-100.0	825/826	99.9%	99.3-100.0
	Frozen	89/90	98.9%	94.0-99.8	992/998	99.4%	98.7-99.7
	Overall	156/157	99.4%	96.5-99.9	1817/1824	99.6%	99.2-99.8
Analyte		TP/(TP+FN)	Sensitivity/ PPA	95% CI	TN/(TN+FP)	Specificity/NPA	95% CI
Influenza Bk	Fresh	64/67	95.5%	87.6 - 98.5	827/828	99.9%	99.3 - 100.0
	Frozen	58/62	93.5%	84.6 - 97.5	1026/1026	100.0%	99.6 - 100.0
	Overall	122/129	94.6%	89.2 - 97.3	1853/1854	99.9%	99.7 - 100.0
Parainfluenza1l	Fresh	3/3	100.0%	43.8 - 100.0	892/892	100.0%	99.6 - 100.0
	Frozen	13/14	92.9%	68.5 - 98.7	1072/1075	99.7%	99.2 - 99.9
	Overall	16/17	94.1%	73.0-99.0	1964/1967	99.8%	99.6-99.9
Parainfluenza 2	Fresh	2/2	100.0%	34.2 - 100.0	893/893	100.0%	99.6 - 100.0
	Frozen	0/0	N/A	N/A	1089/1089	100.0%	99.6 - 100.0
	Overall	2/2	100.0%	34.2 - 100.0	1982/1982	100.0%	99.8 - 100.0
Parainfluenza3m	Fresh	102/104	98.1%	93.3 - 99.5	788/793	99.4%	98.5 - 99.7
	Frozen	9/9	100.0%	70.1 - 100.0	1081/1081	100.0%	99.6 - 100.0
	Overall	111/113	98.2%	93.8 - 99.5	1869/1874	99.7%	99.4 - 99.9
Parainfluenza4n	Fresh	3/3	100.0%	43.8 -100.0	892/892	100.0%	99.6 - 100.0
	Frozen	0/0	N/A	N/A	1087/1089	99.8%	99.3 - 99.9
	Overall	3/3	100.0%	43.8 - 100.0	1979/1981	99.9%	99.6 - 100.0
RespiratorySyncytial Virus(RSV)o	Fresh	73/76	96.0%	88.9 - 98.6	819/820	99.9%	99.3 - 100.0
	Frozen	139/144	96.5%	92.1 - 98.5	941/945	99.6%	98.9 - 99.8
	Overall	212/220	96.3%	93.0 - 98.1	1760/1765	99.7%	99.3 - 99.9
Bacteria
Bordetellapertussisp	Fresh	2/2	100.0%	34.2 - 100.0	893/893	100.0%	99.6 - 100.0
	Frozen	1/1	100.0%	20.7 - 100.0	1082/1088	99.4%	98.8 - 99.7
	Overall	3/3	100.0%	43.8 - 100.0	1975/1981	99.7%	99.3 - 99.9
Chlamydophilapneumoniaeq	Fresh	4/4	100.0%	51.0 - 100.0	891/891	100.0%	99.6 - 100.0
	Frozen	1/1	100.0%	20.7 - 100.0	1087/1088	99.9%	99.5 - 100.0
	Overall	5/5	100.0%	56.6 - 100.0	1978/1979	99.9%	99.7 - 100.0
Mycoplasmapneumoniaer	Fresh	18/18	100.0%	82.4 - 100.0	875/877	99.8%	99.2 - 100.0
	Frozen	1/1	100.0%	20.7 - 100.0	1085/1088	99.7%	99.2 - 99.9
	Overall	19/19	100.0%	83.2- 100.0	1960/1965	99.7%	99.4 - 99.9

Table 5.11: QIAstat-Dx Respiratory Panel prospective clinical performance summarv

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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 during the prospective clinical evaluation. Of these, 95.88% (1912/1994) yielded valid results on the first attempt (i.e., first loaded cartridge). Invalid or no result were obtained for the

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

Conclusions

The QIAstat-Dx Respiratory Panel is substantially equivalent to the legally marketed FilmArray® Respiratory Panel.

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.