The QIAstat-Dx Respiratory Panel Plus is a multiplexed nucleic acid test intended for use with the QIAstat-Dx system for the simultaneous in vitro qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals with clinical signs and symptoms of respiratory tract infection, including SARS-CoV-2.

The following organism types and subtypes are identified using the QIAstat-Dx Respiratory Panel Plus: Adenovirus, Human Coronavirus 229E, Human Coronavirus HKU1, Human Coronavirus NL63, Human Coronavirus OC43, Human Metapneumovirus, Influenza A, Influenza A H1, Influenza A H1N1 pdm09, Influenza B, Parainfluenza virus 1, Parainfluenza virus 2, Parainfluenza virus 3, Parainfluenza virus 4, Respiratory Syncytial Virus, Human Rhinovirus/Enterovirus (not differentiated), Severe Acute Respiratory Syndrome Coronavirus (SARS-COV-2), Bordetella pertussis, Chlamydophila pneumoniae and Mycoplasma pneumoniae.

Nucleic acids from viral and bacterial organisms identified by this test are generally detectable in NPS specimens during the acute phase of infection. Detecting and identifying 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, epidemiological and laboratory findings. The results of this test should not be used as the sole basis for diagnosis, treatment or other patient management decisions.

Negative results in the presence of a respiratory illness may be due to infection with pathogens that are not detected by the test or due to lower respiratory tract infection that is not detected by a NPS specimen.

Conversely, positive results are indicative of the identified microorganism, but do not rule out co-infection with other pathogens not detected by the QIAstat-Dx Respiratory Panel Plus. The agent(s) detected by the QIAstat-Dx Respiratory Panel Plus may not be the definite cause of disease.

The use of additional laboratory testing (e.g., bacterial and viral culture, immunofluorescence and radiography) may be necessary when evaluating a patient with possible respiratory tract infection.

Device Description

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

The QIAstat-Dx Respiratory Panel Plus is intended to be used with I nasopharyngeal swab (NPS) eluted in Universal Transport Media (UTM), which is not provided with the QIAstat-Dx Respiratory Panel Plus.

Once the cartridge has been inserted into the instrument, the test starts automatically and runs for approximately 1 hour. When the test is finished, the cartridge is removed by the user and discarded. The OIAstat-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").

All the reagents required for the complete execution of the test are pre-loaded and selfcontained in the QIAstat-Dx Respiratory Panel Plus cartridge. The user does not need to manipulate any reagents. During the test, reagents are handled by pneumatically-operated microfluidics without any direct contact with the user or the analyzer actuators.

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 (TC) to their intended destinations. Following the introduction of the sample from a disposable transfer pipette, the following assay steps occur automatically and sequentially:

Resuspension of air-dried internal control and Proteinase K (ProtK) enzyme using ● provided buffer and mixing with the liquid sample (IC Cavity and ProtK Cavity);
Cell lysis using mechanical (rotation) and chemical (chaotropic and isotonic) means ● (lysis chamber);
Membrane based nucleic acid purification from Lysate by:
- -Mixing lysate with binding buffer and capturing on the membrane (purification chamber);
- First washing of membrane to remove bound proteins (purification chamber and waste chamber);
- Second washing of membrane to leave only bound nucleic acids -(purification chamber and waste chamber);
- -Rinsing of Transfer Chamber (TC) using the rinsing buffer before introduction of the eluate (Transfer Chamber);
- Drying of membrane with bound nucleic acids with an air flow generated by a high flow vacuum pump (purification chamber);
- -Elution of nucleic acids with elution buffer (purification chamber and TC);
Mixing of the purified nucleic acid (eluate) with lyophilized "Master Mix" reagents (Dry chemistry container (DCC) and TC);
Sequential transfer of defined aliquots of mixed eluate/Master Mix from the . Transfer Chamber to each of eight Reaction Chambers containing the specified, airdried primers and probes;
Within each Reaction Chamber, real-time, multiplex PCR ("rtPCR") testing is . performed. Increase in fluorescence (indicative of detection of each target analyte) is detected directly within each Reaction Chamber;
The detected signal per fluorescent marker per Reaction Chamber is then used by . the system software to generate the assay result.

The QIAstat-Dx Respiratory Panel Plus includes the addition of the SARS-CoV-2 analyte to the analytes that were cleared in the OIAstat-Dx Respiratory Panel (K183597).

AI/ML Overview

The provided text describes the QIAstat-Dx Respiratory Panel Plus, a multiplexed nucleic acid test. The document focuses on demonstrating the substantial equivalence of this new device to a predicate device (BioFire Respiratory Panel 2.1 (RP2.1)), primarily by showing that the new device's SARS-CoV-2 detection is effective and that the addition of SARS-CoV-2 and a change in lysis reagent do not negatively impact the performance of other analytes previously cleared.

Here's an analysis of the acceptance criteria and the study that proves the device meets them, based on the provided text:

Acceptance Criteria and Reported Device Performance

The acceptance criteria are not explicitly stated in a single, concise list with specific thresholds (e.g., "PPA must be >X%"). Instead, they are implicitly demonstrated through performance goals such as high positive percent agreement (PPA) and negative percent agreement (NPA) compared to a comparator method, and meeting detection rates at defined LoD concentrations. For non-SARS-CoV-2 analytes, the criteria seem to be demonstrating equivalence to the previously cleared QIAstat-Dx Respiratory Panel (K183597).

Table of Acceptance Criteria (Implied) and Reported Device Performance

Performance Metric	Implied Acceptance Criteria (Threshold not always explicit, but high % is the goal)	Reported Device Performance	Comments
SARS-CoV-2 Clinical Performance (vs. FDA-cleared RT-PCR comparator)	High Agreement
Positive Percent Agreement (PPA)	High (e.g., >95%)	96.8% (61/63)	One of the two false negative samples was positive by two FDA-EUA molecular SARS-CoV-2 assays (implying the competitor may be better).
Negative Percent Agreement (NPA)	High (e.g., >98%)	99.8% (551/552)	The single false positive sample was positive by two FDA-EUA molecular SARS-CoV-2 assays (implying the 'false positive' was actually a true positive missed by initial comparator).
Representative Panel Equivalency (Non-SARS-CoV-2 Analytes vs. original QIAstat-Dx Respiratory Panel)	High Percent Agreement between the two devices (ideally 100%)
PPA for Influenza B	High	100.0% (20/20)
PPA for Coronavirus OC43	High	100.0% (22/22)
PPA for Parainfluenza virus 3	High	100.0% (24/24)
PPA for Rhinovirus/Enterovirus	High	100.0% (43/43)
PPA for Adenovirus	High	95.0% (38/40)
PPA for Bordetella pertussis	High	100.0% (24/24)
NPA for Influenza B	High	99.4% (167/168)
NPA for Coronavirus OC43	High	100.0% (166/166)
NPA for Parainfluenza virus 3	High	99.4% (163/164)
NPA for Rhinovirus/Enterovirus	High	99.3% (144/145)
NPA for Adenovirus	High	97.3% (144/148)
NPA for Bordetella pertussis	High	99.4% (163/164)
Limit of Detection (LoD) for SARS-CoV-2	≥95% detection rate at LoD concentration
SARS-CoV-2 (USA-WA1-2020)	≥95%	19/20 (95%) at 3160.0 copies/mL
SARS-CoV-2 (England/02/2020)	≥95%	19/20 (95%) at 316.0 copies/mL
SARS-CoV-2 (clinical sample 243)	≥95%	20/20 (100%) at 600.0 copies/mL
SARS-CoV-2 (clinical sample S1229)	≥95%	20/20 (100%) at 1.90E+04 copies/mL
SARS-CoV-2 (clinical sample S1231)	≥95%	20/20 (100%) at 1.90E+04 copies/mL
Analytical Reactivity (Inclusivity) - SARS-CoV-2	100% predicted detection of relevant strains/variants	100% of sequences analyzed predicted to be detected.	Includes Alpha, Beta, Gamma, Lambda, Mu, Delta, Omicron variants.
Analytical Specificity (Exclusivity)	No cross-reactivity with diverse pathogens (beyond noted exceptions)	All on-panel samples generated a positive call for every target and all off-panel targets resulted in negative call, with exception of B. bronchiseptica and B. holmesii cross-reacting with B. pertussis (known for prior device).	In silico analysis also confirmed lack of cross-reactivity for SARS-CoV-2.
Interfering Substances	No inhibition of SARS-CoV-2 detection	None of the tested substances showed inhibition.	Various endogenous, exogenous, and technique-specific substances tested at high concentrations.
Microbial Interference	No inhibition of SARS-CoV-2 detection	All combinations and replicates successfully detected SARS-CoV-2, with minor exceptions needing re-testing or additional strains.
Competitive Inhibition	All targets detected when co-infected	All targets detected.	SARS-CoV-2 at 3x LoD with high concentrations of other on-panel pathogens.
Carryover Study	No carryover between cartridges or chambers	No carryover observed.
Sample Stability	Maintained performance across specified conditions	Performance maintained for Room Temp (4 hrs), Refrigerated (3 days), Frozen (14 days).
Precision	High detection rates at 1x LoD and 3x LoD concentrations, and 100% for negative samples (low variability)	(See Table 5.9 for detailed percentages per analyte)	Demonstrated robust performance across variables (days, operators, instruments, lots). Confirmed acceptance criteria met.

Study Details:

Sample Size Used for the Test Set and Data Provenance:
- SARS-CoV-2 Clinical Performance: 616 prospective NPS specimens (615 included in analysis due to one exclusion).
  - Provenance: Collected from "five (5) geographically diverse study sites in the U.S.", indicating prospective data from multiple locations within the US. Collection period: February 2023 to May 2023 and during February 2024.
- Representative Panel Equivalency (Non-SARS-CoV-2): 190 de-identified clinical NPS specimens.
  - Provenance: Clinical NPS specimens, both positive and negative as per Standard of Care. No specific country mentioned, but likely US given the overall context of FDA submission. "De-identified clinical NPS specimens" suggests retrospective collection but could also be prospective if de-identified at collection.
Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts:
- Clinical Performance (SARS-CoV-2): For SARS-CoV-2, the ground truth was established by comparing to "an FDA-cleared SARS-CoV-2 RT-PCR comparator method." This implies an established and validated laboratory method, not human expert consensus, for determining the presence or absence of the virus.
- Representative Panel Equivalency: For the representative panel analytes, the comparison was against "Standard of Care" and by parallel testing with the "QIAstat-Dx Respiratory Panel" (the predicate device for these analytes). Again, this points to laboratory reference methods rather than human expert interpretation of imaging/clinical findings.
- The document does not mention the use of human experts (e.g., radiologists) for establishing ground truth, as the device is an in-vitro diagnostic (IVD) for molecular detection, not an imaging AI.
Adjudication Method for the Test Set:
- Clinical Performance (SARS-CoV-2): For discrepant results with the primary comparator for SARS-CoV-2, the text mentions that the "two samples with false negative SARS-CoV-2 results by the QIAstat-Dx Respiratory Panel Plus were both positive by two FDA-EUA molecular SARS-CoV-2 assays." Similarly, the "single sample with a false positive SARS-CoV-2 result...was positive by two FDA-EUA molecular SARS-CoV-2 assays." This indicates a form of adjudication by a secondary, independent highly sensitive molecular test (two FDA-EUA assays) to resolve discrepancies, providing a more robust ground truth.
- Representative Panel Equivalency: No specific adjudication method is mentioned for this section beyond the direct comparison to "Standard of Care" and the QIAstat-Dx Respiratory Panel.
If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done:
- No. An MRMC study is relevant for imaging devices where human readers interpret images. This device is an in-vitro diagnostic (IVD) for nucleic acid detection, performed automatically by an instrument. There are no human readers "interpreting" the output in a way that would necessitate an MRMC study.
If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done:
- Yes, the performance data (PPA, NPA, LoD, Analytical Reactivity, Specificity, Interference, Precision) are all reported as standalone performance of the QIAstat-Dx system. The system automatically interprets test results and displays a summary (page 6). This is an "algorithm only" performance in the context of an automated IVD test.
The Type of Ground Truth Used:
- Lab-based Comparator Methods: For SARS-CoV-2 clinical performance, the ground truth was established using an "FDA-cleared SARS-CoV-2 RT-PCR comparator method" with further confirmation by "two FDA-EUA molecular SARS-CoV-2 assays" for discrepant results.
- For the non-SARS-CoV-2 analytes, ground truth was derived from "Standard of Care" lab results and comparison to the "QIAstat-Dx Respiratory Panel" (the previously cleared device).
- For analytical studies (LoD, inclusivity, specificity, etc.), ground truth was established by precise laboratory spiking of known concentrations of pathogens or interferents.
- This is molecular/pathology-based ground truth, not outcomes data or expert consensus based on clinical or imaging findings.
The Sample Size for the Training Set:
- The document does not provide information on the training set for the QIAstat-Dx Respiratory Panel Plus. This document is a 510(k) summary, which focuses on validation and demonstrated substantial equivalence to a predicate device. Information about the training data for the underlying assays or software algorithms is typically part of the device's development and design control documentation, which is not usually disclosed in a public 510(k) summary. The "device performance" described here refers to the testing of the final, developed device.
How the Ground Truth for the Training Set Was Established:
- As the training set information is not provided, how its ground truth was established is also not detailed. In general, for molecular diagnostics like this, training data (if any specific to machine learning/AI components were used in the assay development) would typically involve well-characterized clinical samples or contrived samples with known pathogen presence/absence and concentration, verified by highly sensitive and specific reference methods (e.g., Sanger sequencing, quantitative PCR).

Summary

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May 10, 2024

Image /page/0/Picture/1 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left side of the logo is the Department of Health & Human Services logo. To the right of that is a blue square with the letters FDA in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

QIAGEN GmbH % Melissa Mahall Senior Director, Regulatory Affairs Oiagen 19300 Germantown Road Germantown, Maryland 20874

Re: K233100

Trade/Device Name: QIAstat-Dx Respiratory Panel Plus Regulation Number: 21 CFR 866.3981 Regulation Name: Device To Detect And Identify Nucleic Acid Targets In Respiratory Specimens From Microbial Agents That Cause The SARS-Cov-2 Respiratory Infection And Other Microbial Agents When In A Multi-Target Test Regulatory Class: Class II Product Code: QOF Dated: September 26, 2023 Received: September 26, 2023

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 (the 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 available 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.

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Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30. Design controls; 21 CFR 820.90. Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).

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 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 Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). 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 (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Joseph Briggs -S

Joseph Briggs, Ph.D. Deputy Branch Chief Viral Respiratory and HPV Branch Division of Microbiology Devices

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OHT7: Office of In Vitro Diagnostics 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) K233100

Device Name QIAstat-Dx® Respiratory Panel Plus

Indications for Use (Describe)

The use of additional laboratory testing (e.g., bacterial and viral culture, immunofluorescence and radiography) may be necessary when evaluating a patient with possible respiratory tract infection.

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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510(k) Summary

General Information

Submitted by:	QIAGEN GmbHQIAGEN Strasse 1Hilden, Germany 40724
Contact Person:	Melissa MahallSenior Director, Regulatory AffairsQIAGEN19300 Germantown RoadGermantown, MD 20874Phone: 301-944-7768Email: melissa.mahall@qiagen.com
Date Prepared:	May 08, 2024
Device Name:	QIAstat-Dx Respiratory Panel Plus
Classification:	21 CFR 866.3981 - Device To Detect And Identify Nucleic AcidTargets In Respiratory Specimens From Microbial Agents ThatCause The SARS-Cov-2 Respiratory Infection And OtherMicrobial Agents When In A Multi-Target Test
Product Code:	QOF
Predicate Device:

Manufacturer	Product Name	De Novo/510(k) No.
BioFire Diagnostics, LLC	BioFire Respiratory Panel 2.1 (RP2.1)	DEN200031

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Device Description

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

Resuspension of air-dried internal control and Proteinase K (ProtK) enzyme using ● provided buffer and mixing with the liquid sample (IC Cavity and ProtK Cavity);
Cell lysis using mechanical (rotation) and chemical (chaotropic and isotonic) means ● (lysis chamber);
Membrane based nucleic acid purification from Lysate by:
- -Mixing lysate with binding buffer and capturing on the membrane (purification chamber);
- First washing of membrane to remove bound proteins (purification chamber and waste chamber);
- Second washing of membrane to leave only bound nucleic acids -(purification chamber and waste chamber);
- -Rinsing of Transfer Chamber (TC) using the rinsing buffer before introduction of the eluate (Transfer Chamber);
- Drying of membrane with bound nucleic acids with an air flow generated by a high flow vacuum pump (purification chamber);
- -Elution of nucleic acids with elution buffer (purification chamber and TC);

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Mixing of the purified nucleic acid (eluate) with lyophilized "Master Mix" reagents (Dry chemistry container (DCC) and TC);
Sequential transfer of defined aliquots of mixed eluate/Master Mix from the . Transfer Chamber to each of eight Reaction Chambers containing the specified, airdried primers and probes;
Within each Reaction Chamber, real-time, multiplex PCR ("rtPCR") testing is . performed. Increase in fluorescence (indicative of detection of each target analyte) is detected directly within each Reaction Chamber;
The detected signal per fluorescent marker per Reaction Chamber is then used by . the system software to generate the assay result.

The QIAstat-Dx Respiratory Panel Plus includes the addition of the SARS-CoV-2 analyte to the analytes that were cleared in the OIAstat-Dx Respiratory Panel (K183597).

Intended Use

The following organism types and subtypes are identified using the QIAstat-Dx Respiratory Panel Plus: Adenovirus, Human Coronavirus 229E, Human Coronavirus HKU1, Human Coronavirus NL63, Human Coronavirus OC43, Human Metapneumovirus, Influenza A, Influenza A H1, Influenza A H1N1 pdm09, Influenza A H3, Influenza B, Parainfluenza virus 1, Parainfluenza virus 2, Parainfluenza virus 3, Parainfluenza virus 4, Respiratory Syncytial Virus, Human Rhinovirus/Enterovirus (not differentiated), Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2), Bordetella pertussis, Chlamydophila pneumoniae and Mycoplasma pneumoniae.

Conversely, positive results are indicative of the presence of the identified microorganism, but do not rule out co-infection with other pathogens not detected by the QIAstat-Dx

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Respiratory Panel Plus. The agent(s) detected by the QIAstat-Dx Respiratory Panel Plus may not be the definite cause of disease.

The use of additional laboratory testing (e.g., bacterial and viral culture, immunofluorescence and radiography) may be necessary when evaluating a patient with possible respiratory tract infection.

Comparison of the QIAstat-Dx Respiratory Panel Plus and the Predicate Device

Similarities and differences between the QIAstat-Dx Respiratory Panel Plus and the predicate device are shown in Table 5.1.

	Table 5.1: Comparison of the QIAstat-Dx Respiratory Panel Plus with the predicate
device
Characteristic	Subject Device	Predicate
Name	QIAstat-Dx Respiratory Panel Plus	BioFire Respiratory Panel 2.1 (RP2.1)
De Novo/510(k) No.	K233100	DEN200031
Regulation	21 CFR 866.3981	21 CFR 866.3981
Product Code	QOF	QOF
Device Class	Class II	Class II
Similarities
Intended Use	The QIAstat-Dx Respiratory Panel Plus is a multiplexed nucleic acid test intended for use with the QIAstat-Dx system for the simultaneous in vitro qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals with clinical signs and symptoms of respiratory tract infection, including SARS-CoV-2.	The BioFire Respiratory Panel 2.1 (RP2.1) is a PCR-based multiplexed nucleic acid test intended for use with the BioFire FilmArray 2.0 or BioFire FilmArray Torch systems for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections, including COVID-19.
	The following organism types and subtypes are identified using the QIAstat-Dx Respiratory Panel Plus:Adenovirus,Coronavirus 229E, HumanCoronavirus HKU1, Human	The following organism types and subtypes are identified using the BioFire RP2.1:• Adenovirus,• Coronavirus 229E,• Coronavirus HKU1,
Characteristic	Subject Device	Predicate
	Coronavirus NL63, HumanCoronavirus OC43, HumanMetapneumovirus, Influenza A, Influenza A H1, Influenza AH1N1 pdm09, Influenza A H3,Influenza B, Parainfluenzavirus 1, Parainfluenza virus 2,Parainfluenza virus 3,Parainfluenza virus 4,Respiratory Syncytial Virus,Human Rhinovirus/Enterovirus(not differentiated), SevereAcute Respiratory SyndromeCoronavirus (SARS-CoV-2),Bordetella pertussis,Chlamydophila pneumoniaeand Mycoplasma pneumoniae.	• Coronavirus NL63,• Coronavirus OC43,• Severe AcuteRespiratory SyndromeCoronavirus (SARS-CoV-2),• Human Metapneumovirus,• Human Rhinovirus/Enterovirus,• Influenza A, including subtypes H1, H1-2009, and H3,• Influenza B,• Parainfluenza Virus 1,• Parainfluenza Virus 2,• Parainfluenza Virus 3,• Parainfluenza Virus 4,• Respiratory Syncytial Virus,• Bordetella parapertussis (IS1001),• Bordetella pertussis (ptxP),• Chlamydia pneumoniae, and• Mycoplasma pneumoniae
	Nucleic acids from viral andbacterial organisms identifiedby this test are generallydetectable in NPS specimensduring the acute phase ofinfection. Detecting andidentifying specific viral andbacterial nucleic acids fromindividuals presenting withsigns and symptoms of arespiratory infection aids in thediagnosis of respiratoryinfection, if used in conjunctionwith other clinical,epidemiological and laboratoryfindings. The results of this testshould not be used as the solebasis for diagnosis, treatment orother patient managementdecisions.Negative results in the presenceof a respiratory illness may bedue to infection with pathogensthat are not detected by the test	Nucleic acids from therespiratory viral and bacterialorganisms identified by this testare generally detectable in NPSspecimens during the acutephase of infection. The detectionand identification of specificviral and bacterial nucleic acidsfrom individuals exhibiting signsand/or symptoms of respiratoryinfection is indicative of thepresence of the identifiedmicroorganism and aids in thediagnosis of respiratory infection
Characteristic	Subject Device	Predicate
	infection that is not detected bya NPS specimen.	clinical and epidemiologicalinformation. The results of thistest should not be used as the solebasis for diagnosis, treatment, orother patient managementdecisions.
	Conversely, positive results areindicative of the presence of theidentified microorganism, butdo not rule out co-infectionwith other pathogens notdetected by the QIAstat-DxRespiratory Panel Plus. Theagent(s) detected by theQIAstat-Dx Respiratory PanelPlus may not be the definitecause of disease.	Negative results in the setting ofa respiratory illness may be dueto infection with pathogens thatare not detected by this test, orlower respiratory tract infectionthat may not be detected by anNPS specimen. Positive resultsdo not rule out coinfection withother organisms. The agent(s)detected by the BioFire RP2.1may not be the definite cause ofdisease. Additional laboratorytesting (e.g. bacterial and viralculture, immunofluorescence,and radiography) may benecessary when evaluating apatient with possible respiratorytract infection.
	The use of additionallaboratory testing (e.g.,bacterial and viral culture,immunofluorescence andradiography) may be necessarywhen evaluating a patient withpossible respiratory tractinfection.
Specimen Type	Nasopharyngeal swabs (NPS)	Nasopharyngeal swabs (NPS)
Organism Detected	See analyte list above,RNA/DNA	See analyte list above,RNA/DNA
Amplification andDetectionTechnology	PCR	PCR

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Characteristic	Subject Device	Predicate
Assay Controls	One internal control in each cartridge to control for sample processing that is subjected to all nucleic acid extraction and amplification steps similar to patient samples. Instructions for Use indicates quality control requirements should be performed in conformance with local, state, and/or federal regulations or accreditation requirements and the laboratory's standard quality control procedures.	Two controls are included in each reagent pouch to control for sample processing and both stages of PCR and melt analysis. Labeling recommends the use of external positive and negative controls regularly. Use viral transport medium as the external negative control, and previously characterized positive samples or negative samples spiked with well characterized organisms as external positive controls.
	Differences
	Characteristic	Subject Device	Predicate
	Bordetella Assay Targets	The QIAstat-Dx Respiratory Panel Plus detects Bordetella pertussis.	The BioFire Respiratory Panel 2.1 (RP2.1) detects Bordetella pertussis (ptxP) and Bordetella parapertussis (IS1001).
	Nucleic Acid Extraction	Extraction of nucleic acids using spin columns	Extraction of nucleic acids using magnetic beads
	Detection Technology	Detection of amplified targets uses an increase in fluorescence to generate the assay results.	Melting curve analysis is used to confirm the identity of amplified targets to generate assay results.
	Operational	The sample is loaded straight into the cartridge.	The pouch is hydrated prior to introducing the sample.
	Amplification and Detection Instrument	QIAstat-Dx Analyzer 1.0	FilmArray 2.0 or FilmArray

Performance Characteristics - Clinical Studies

SARS-CoV-2

The clinical performance of the SARS-CoV-2 target in the QIAstat-Dx Respiratory Panel Plus was established through studies conducted at five (5) geographically diverse study sites in the U.S. The sites were representative of testing environments where the device will ultimately be used. The testing was performed by laboratory personnel likely to perform the testing in clinical practice after device marketing. Nasopharyngeal swab (NPS) specimens were prospectively collected from patients with signs and symptoms of respiratory infection.

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A total of 616 prospective NPS specimens in UTM were enrolled and tested in a comparison study. One specimen was excluded due to not meeting the inclusion criteria, so 615 specimens were included in the analysis. From February 2023 to May 2023 and during February 2024, NPS specimens were prospectively collected from individuals meeting the study inclusion criteria. NPS specimens were tested fresh on both the QIAstat-Dx Respiratory Panel Plus and an FDA-cleared SARS-CoV-2 RT-PCR comparator method.

The performance of the SARS-CoV-2 target in the OlAstat-Dx Respiratory Panel Plus was evaluated by comparing the SARS-CoV-2 QIAstat-Dx Respiratory Panel Plus result with the result from an FDA-cleared SARS-CoV-2 RT-PCR comparator method. The SARS-CoV-2 QIAstat-Dx Respiratory Panel Plus prospective performance data in positive percent and negative percent against the comparator method are presented in Table 5.2.

Table 5.2: QIAstat-Dx Respiratory Panel Plus SARS-CoV-2 prospective clinical performance summary

	Positive Percent Agreement		Negative Percent Agreement
Target	TP/(TP+FN)	%	TN/(TN+FP)	%	95% CI
SARS-CoV-2	61 / 631	96.8%	551 / 5522	99.8%	89.0%-99.6%
					99.0%-100.0%

TP-True Positive, FP-False Positive, TN-True Negative, FN-False Negative

1The two samples with false negative SARS-CoV-2 results by the QIAstat-Dx Respiratory Panel Plus were both positive by two FDA-EUA molecular SARS-CoV-2 assays.

2The single sample with a false positive SARS-CoV-2 result by the QIAstat-Dx Respiratory Panel Plus was positive by two FDA-EUA molecular SARS-CoV-2 assays.

There were two specimens that yielded a co-infection including SARS-CoV-2 by the QIAstat-Dx Respiratory Panel Plus. One was positive for Human Metapneumovirus and the second was positive for Rhinovirus/Enterovirus. Both results were true positive based on the comparator result.

Representative Panel Equivalency

Clinical performance for the non-SARS-CoV-2 analytes were established previously in K183597. To confirm the clinical performance of the original panel analytes and to demonstrate clinical performance equivalence between the two products (the previously cleared QIAstat-Dx Respiratory Panel in K183597 and QIAstat-Dx Respiratory Panel Plus) a subset of original panel analytes were chosen as a representative panel for an internal comparison study. The representative panel covers all target types (RNA, DNA, Bacteria, Viral), one target from each Reaction Chamber within the test cartridge and was chosen based on the prevalence of the pathogen, including:

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1. Influenza B
1. Coronavirus OC43
1. Parainfluenza virus 3
1. Rhinovirus/enterovirus
1. Adenovirus
1. Bordetella pertussis

A total of 190 de-identified clinical NPS specimens, both positive and negative as per Standard of Care, were obtained and tested for the representative panel analytes. Each specimen was run in parallel on QIAstat-Dx Respiratory Panel and QIAstat-Dx Respiratory Panel Plus. The performance data in positive percent and negative percent agreements are summarized in Table 5.3.

Grouping Variable(s)		Proportiona		Two-Sided 95% Confidence Limit
Analysis	Pathogen	Fraction	Percentage	Lower	Upper
PPA	Influenza B	20 / 20	100.0%	83.2%	100.0%
	Coronavirus OC43	22 / 22	100.0%	84.6%	100.0%
	Parainfluenza virus 3	24 / 24	100.0%	85.8%	100.0%
	Rhinovirus / Enterovirus	43 / 43	100.0%	91.8%	100.0%
	Adenovirus	38 / 40	95.0%	83.1%	99.4%
	Bordetella pertussis	24 / 24	100.0%	85.8%	100.0%
NPA	Influenza B	167 / 168	99.4%	96.7%	100.0%
	Coronavirus OC43	166 / 166	100.0%	97.8%	100.0%
	Parainfluenza virus 3	163 / 164	99.4%	96.7%	100.0%
	Rhinovirus / Enterovirus	144 / 145	99.3%	96.2%	100.0%
	Adenovirus	144 / 148	97.3%	93.2%	99.3%
	Bordetella pertussis	163 / 164	99.4%	96.7%	100.0%

Table 5.3: PPA and NPA of the Representative Panel analytes between QIAstat-Dx Respiratory Panel Plus and QIAstat-Dx Respiratory Panel

a The performance of the QlAstat-Dx Respiratory Panel Plus was established by comparing to results from the QIAstat-Dx Respiratory Panel.

Clinical performance equivalence between the two products (QIAstat-Dx Respiratory Panel and QIAstat-Dx Respiratory Panel Plus) has been demonstrated showing that the addition of SARS-CoV-2 and change in the lysis reagent had no impact to the performance of the original panel analytes.

Performance Characteristics - Non-clinical Studies

The studies presented have been performed to demonstrate the non-clinical performance of the SARS-CoV-2 assay. In addition, where indicated, a subset of original panel analytes were tested to demonstrate the addition of SARS-CoV-2 and change in the lysis reagent does not impact the performance of the original panel analytes. The performance of the original panel analytes is described in K183597.

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Limit of Detection

The Limit of Detection (LoD) is defined as the lowest concentration at which ≥95% of the tested sample generates a positive call. A total of 5 SARS-CoV-2 strains were evaluated in the LoD study by analyzing serial dilutions of contrived samples prepared from culture isolates obtained from commercial suppliers or clinical samples positive for the target analyte. Samples were prepared by spiking SARS-CoV-2 into NPS matrix. The concentration estimated to be the LoD was confirmed by testing 20 replicates during at least 3 different days by different operators using at least four different lots of QIAstat-Dx Respiratory Panel Plus cartridges executed on 3 or more QIAstat-Dx Analyzer 1.0 instruments.

Individual LoD concentrations for each SARS-CoV-2 strain is shown in Table 5.4.

Pathogen	Source/Catalog ID	Strain	LoD concentration Detection(copies/mLª)	rate
SARS-CoV-2	ZeptoMetrix/0810587CFH	USA-WA1-2020	3160.0	19/20
SARS-CoV-2	NIBSC/First WHO InternationalStandard for SARS-CoV-2RNA 20/146	England/02/2020	316.0	19/20
SARS-CoV-2	STAT-Dx Life, S.L. -Clinical sample/243	n/a	600.0	20/20
SARS-CoV-2	Vall d'Hebron hospital -Clinical sample/S1229	n/a	1.90E+04	20/20
SARS-CoV-2	Vall d'Hebron hospital -Clinical sample/S1231	n/a	1.90E+04	20/20

Table 5.4: LoD concentrations for SARS-CoV-2 strains tested with QIAstat-Dx Respiratory Panel Plus

a Titer determined in molecular units (copies/mL) by in-house developed and validated qPCR assay.

LoD and equivalency for the previously cleared QIAstat-Dx Respiratory Panel cartridges and the new QIAstat-Dx Respiratory Panel Plus cartridges were confirmed by testing a subset of original panel analytes. Testing consisted of twenty replicates tested side-by-side on the cleared device and the new device containing SARS-CoV-2 at the LoD concentration as defined for the cleared device (QIAstat-Dx Respiratory Panel) verifying that the addition of the SARS-CoV-2 assay and change in the lysis reagent does not impact the performance of targets in other reaction chambers.

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Analytical Reactivity (Inclusivity)

QIAGEN routinely evaluates the overall inclusivity of the entire set of SARS-CoV-2 available genomes, including all relevant variants and lineages. This includes (1) Inclusivity Analysis and mismatch detection among SARS-CoV-2 strains and (2) same analysis focused on specific SARS-CoV-2 variants or lineages, evaluating the possible effect of detected mismatches in the QIAstat-Dx performance. Only critical mismatches established were evaluated in the laboratory.

In total. 8.118.241 available genomes since 01.01.2020 until 06.05.2022 from around the globe were analyzed for inclusivity (additional vigilance of SARS-CoV-2 genomic variability is continuously done internally in the Post-Market Surveillance OIAGEN monitoring study). Of them, 7,932,071 (97.71%) presented no evidence of mismatches in the targeted regions. The remainder of genomes (2.01%) presented any mismatch among the sequence in the binding region, but only 19,045 (0.23%) presented a mismatch (3'-end of any primer) in a critical position and frequency >0.2%. Genetic patterns included among those sequences were evaluated in laboratory testing, with no loss of performance at LoD level. Consequently, 100% of sequences analyzed were predicted to be detected.

Variants of Concern, of Interest and Under Investigation (VOCs, VOIs, VUI respectively) were also evaluated from the 8,118,241 sequences. After classifying all previously analyzed genomes into lineages based on GISAID and PANGO classifications, the following variants were included: Alpha (9 clades), Beta (5 clades), Gamma (23 clades), Lambda (2 clades), Mu (4 clades), Delta (242 clades), Omicron (73 clades). As a result, all variants and lineages described are 100% predicted to be detected.

For the rest of the analytes detected by the QIAstat-Dx Respiratory Panel Plus, an updated in silico analysis was performed to confirm previous device inclusivity results. All single primers and probes corresponding to targets included in the QIAstat-Dx Respiratory Panel Plus (non-SARS-CoV-2 targets) were analyzed to detect the specificity of the Panel using BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi).

All QIAstat-Dx Respiratory Panel Plus primers and probes are predicted to be inclusive for all clinical prevalent and relevant strains for each pathogen against all available sequences in the NCBI data base.

Analytical Specificity (Exclusivity)

The potential for cross-reactivity between all QIAstat-Dx Respiratory Panel Plus on-panel organisms (including SARS-CoV-2) and various on-panel or off-panel organisms that can happen in clinical respiratory specimens was evaluated with a combination of laboratory (in vitro) testing and in silico characterization. For laboratory testing, on-panel (intra-panel cross-reactivity) and off-panel tested organisms (not covered by the panel content and, therefore, not intended to be detected by the QIAstat-Dx Respiratory Panel Plus) complement the organisms evaluated in the previously cleared device (OIAstat-Dx Respiratory Panel) and add additional organisms selected to assess the specificity of the

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new SARS-CoV-2 assays included in the Panel. Additionally, all single primers and probes for the SARS-CoV-2 detection were analyzed in silico to complement the overall specificity prediction of the device. On-panel organisms were tested to assess the potential for intra-panel cross-reactivity and off-panel organisms were tested to evaluate panel exclusivity. These organisms included those which are related to, but distinct from, respiratory panel organisms or that could be present in specimens collected from the intended test population. Selected organisms are clinically relevant (colonizing the upper respiratory tract or causing respiratory symptoms), are common skin flora or laboratory contaminants, or are microorganisms by which much of the population may have been infected. Both on-panel and off-panel organisms tested are shown in Table 5.5.

Samples were prepared by spiking potential cross-reactive organisms into simulated nasopharyngeal swab sample matrix at the highest concentration possible based on the organism stock, preferably 105 TCID50/mL for viral targets and 106 CFU/mL for bacterial and fungal targets. All on-panel samples generated a positive call for every target and all off-panel targets resulted in negative call, with the exception of B. bronchiseptica and B. holmesii with a cross-reaction with B. pertussis. Those cross-reactions were described in the cleared device, and they are expected to not have clinical relevance since the presence of other Bordetella species are usually associated as a co-infection with Bordetella pertussis.

On-Panel	Pathogen	Cross-reactivity detected
Bacteria
	Mycoplasma pneumoniae	None
	Bordetella pertussisc	None
	Chlamydophila pneumoniae	None
Viruses
	Influenza A H1N1c	None
	Influenza A H3N2	None
	Influenza A/2009/H1N1	None
	Influenza B	None
	Human Coronavirus 229Ec	None
	Human Coronavirus OC43c	None
	Human Coronavirus NL63	None
	Human Coronavirus HKU1	None
	Parainfluenza virus 1	None
	Parainfluenza virus 2	None
	Parainfluenza virus 3c	None
	Parainfluenza virus 4	None
	RSV Ac	None
	hMPVc	None
	Adenovirus A12c	None
Pathogen	Cross-reactivity detected
Adenovirus C	None
Adenovirus B	None
Enterovirus D68c	None
Rhinovirusc	None
Echovirus 6c	None
SARS-CoV-2	None
Off-Panel
Bacteria
Acinetobacter calcoaceticus	None
Bordetella avium	None
Bordetella bronchiseptica	Bordetella pertussisa
Bordetella hinzii	None
Bordetella holmesii	Bordetella pertussisa
Corynebacterium diphteriaec	None
Enterobacter aerogenes	None
Haemophilus influenzaec	None
Klebsiella oxytoca	None
Lactobacillus acidophilus	None
Legionella feeleii	None
Legionella micdadei	None
Legionella pneumophilac	None
Mycobacterium tuberculosis	None
Mycoplasma genitalium	None
Mycoplasma hominis	Noneb
Mycoplasma orale	None
Neisseria elongata	None
Neisseria gonorrhoeae	None
Serratia marcescens	None
Staphylococcus aureusc	None
Staphylococcus epidermidis	None
Stenotrophomonas maltophilia/Pseudomonas maltophilia	None
Streptococcus pneumoniaec	None
Streptococcus salivariusc	None
Ureaplasma urealyticum	None
Escherichia coli (0157)	None
Klebsiella pneumoniae	None
Moraxella catarrhalis /Branhamella catarrhalis	None
Neisseria meningitidisc	None
Proteus mirabilis	None
Pseudomonas aeruginosac	None
Pathogen	Cross-reactivity detected
Streptococcus pyogenes	Noneb
Streptococcus agalactiae	None
Viruses
Bocavirus	None
Cytomegalovirusc	None
Epstein-Barr Virus	None
Measles Virus	None
MERS Coronavirus	None
Mumps	None
Severe Acute Respiratory Syndrome (SARS) virus	None
Herpes Simplex Virus 1c	None
Herpes Simplex Virus 2	None
Fungi
Candida albicansc	None
Aspergillus flavus	None
Aspergillus fumigatus	Noneb
Cryptococcus neoformans	None

Table 5.5: List of Analytical Specificity Pathogens

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a. As described for the previously cleared device, targeted gene for Bordetella pertussis (IS481) is a mobile transposon also present in B. holmesii (detected in 6/6 replicates) and some clusters of B. bronchiseptica (detected in 3/3 replicates), so observed cross-reaction with B. pertussis was expected in both cases.

b. B. pertussis was detected with high Ct value (low concentration) for A. fumigatus (1/6 replicates). Mycoplasma hominis (1/9 replicates) and S. pyogenes (2/6 replicates). Proper investigation confirmed B. pertussis contamination in all cases. In silico analysis confirmed lack of crossreactivity of B. pertussis assay with all three organisms.
c. Pathogen tested in combination with SARS-CoV-2 at 3xLoD resulting in no impact on assay performance.

Since the QIAstat-Dx Respiratory Panel Plus differs only from the previously cleared device (QIAstat-Dx Respiratory Panel) by the addition of primers and probes for SARS-CoV-2 detection and a change in the lysis reagent, in silico evaluation of the two SARS-CoV-2 assays was performed to corroborate lack of cross-reactivity of the device. An exclusivity BLAST-based in silico screening was performed on week 16, 2022 (April 20th 2022), including all available sequences in GenBank database. No GenBank sequences corresponding to human-infective pathogens were predicted to be positive for the primer/probe sets included in the QIAstat-Dx system to detect SARS-CoV-2.

Interfering Substances

Potential Interfering substances (endogenous, exogenous and technique-specific) were tested at a concentration predicted to be above the levels expected to be found in an authentic NPS specimen for the SARS-CoV-2 target. Interfering substances for all the other targets of the QIAstat-Dx Respiratory Panel Plus have been tested using the cleared device (QIAstat-Dx Respiratory Panel) with the results remaining applicable, as the only

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differences between the devices are the addition of SARS-CoV-2 analyte and a change in the lysis reagent .

None of the substances tested showed inhibition of SARS-CoV-2 (see Table 5.6).

Substance Type	Substance / Pathogen	Concentration tested
EndogenousSubstances	Human genomic DNA 200 ng/µL	20 ng/µL
	Human Blood (+NaCitrate)	1% v/v
	Mucin from bovine submaxillary glands	1% v/v
	Tobramycin (systemic antibiotic)	0.6 mg/ml
	Mupirocin	2% w/v
	OCEAN Saline nasal spray	1% v/v
Exogenoussubstances	Afrin®, severe congestion nasal spray(Oxymetazoline HCl)	1% v/v
	Analgesic ointment (Vicks® VapoRub®)	1% w/v
	Petroleum Jelly (Vaseline®)	1% w/v
	Chiroflu Influenza Vaccine (surface antigeninactivated)	0.000001% v/v
	Disinfecting wipes	1/2 inches2/1 ml UTM
	DNAZap	1% v/v
	ProtectRNATM RNase Inhibitor 500x Concentrate	1% v/v
	Bleach	5% v/v
	Ethanol	5% v/v
TechniqueSpecificSubstances	Swab Copan 168C	1 swab/1 ml UTM
	Swab Copan FloQ	1 swab/1 ml UTM
	Swab Copan 175KS01	1 swab/1 ml UTM
	Swab Puritan 25-801 A 50	1 swab/1 ml UTM
	VTM Sigma Virocult	100%
	VTM Remel® M4-RT	100%
	VTM RT	100%
	BD Universal Viral Transport	100%
	DeltaSwab Virus	100%

Table 5.6: Potential Interfering Substances Concentration without observable inhibitory effect on SARS-CoV-2

Microbial Interference

A microbial interference study was conducted to assess the inhibitory effects of select nontarget organisms on the ability to detect SARS-CoV-2. Clinically relevant and challenging concentrations (1.00E+06 CFU/mL for bacteria/fungi, 1.00E+05 PFU/mL for viruses unless otherwise noted) of non-target organisms were individually mixed with SARS-CoV-2 at 3x LoD in simulated NPS matrix. Testing was performed in triplicate with two additional tests performed if SARS-CoV-2 was not detected in one of the original three

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replicates. All combinations and replicates successfully detected SARS-CoV-2 except for three samples, one Legionella pneumophila, one Streptococcus salivarius sample, and one H.influenzae sample. For these, additional replicates successfully detected SARS-CoV-2. Where available, at least one additional strain of L. pneumophila, S. salivarius or H.influenzae was also tested in triplicate with all samples successfully detecting SARS-CoV-2. See Table 5.7 for a list of the strains tested and the result summary.

Non-Target Organism	Strain/Isolate	Source/ Catalog #	# SARS-CoV-2detected/ validruns
Staphylococcus aureusa	FDA 209	ATCC CRM-6538	3/3
Streptococcus pneumoniae	Z022 19F	ZeptoMetrix 0801439	3/3
Streptococcus salivarius	C699 [S30D]	ATCC 13419	3/3
Streptococcus salivarius	Z127	ZeptoMetrix 0801896	4/5
Haemophilus influenzae	AMC 36-A-7	ATCC 8142	3/3
Haemophilus influenzae	AMC 36-A-1	ATCC 10211	3/3
Candida albicans	CBS 562	ATCC 18804	3/3
Herpes Simplex Virus 1	ATCC-2011-9	ATCC VR-1789	3/3
Staphylococcus epidermidis	Fussel	ATCC 14990	4/5
Pseudomonas aeruginosa	PRD-10	ATCC 15442	3/3
Legionella pneumophila	Philadelphiab	ZeptoMetrix 0801645	3/5
Legionella pneumophila	Philadelphia-1b	ATCC 33152	3/3
Legionella pneumophila	Los Angeles-1	ATCC 33156	3/3
Neisseria meningitidisa	serogroup A	ATCC 13077	3/3
Corynebacteriumdiphtheriaea	48255	ATCC 11913	3/3
Human Cytomegalovirus(CMV)a	Towne	Zeptometrix 0810499CFHI	3/3

Table 5.7: Microbial Interference Study Results

a S. aureus evaluated at 4.5x108 CFU/mL, N. meningitidis 1.0x103 CFU/mL, C. diphtheriae 1.0x103 CFU/mL, and CMV at 1.0x104 TCID50/mL.

b Philadelphia and Philadelphia-1 are both designations of strain Philadelphia serogroup-1, differences in naming are due to supplier.

Competitive Inhibition

Clinically relevant co-infections testing demonstrated that when at least two QIAstat-Dx Respiratory Panel Plus pathogens of different concentrations are simultaneously present in one sample all targets can be detected by the assay. SARS-CoV-2 at 3x LoD has been tested in combination with the following on-panel pathogens at high concentrations (10E+05 PFU/mL for viral targets, 10E+06 CFU/mL for bacterial targets), with no impact on assay performance: Coronavirus 229E, Coronavirus OC43, Adenovirus A12, Parainfluenza Virus 3, Bordetella pertussis, Enterovirus D68, Echovirus 6, Respiratory Syncytial Virus (RSV), Rhinovirus, hMPV and Influenza A H1N1.

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Carryover Study

A carryover study was performed using the QIAstat-Dx Respiratory Panel Plus to evaluate the potential occurrence of cross-contamination between consecutive runs and carryover between cartridge chambers on the QIAstat-Dx Analyzer 1.0. Two panels consisting of high concentrations of influenza A, Parainfluenza virus 3, and M. pneumoniae (panel A) or Coronavirus NL63, RSV, and SARS-CoV-2 (panel B) in simulated NPS matrix were each tested multiple times between runs of negative (no analyte) cartridges for a total of 62 runs across several days. No carryover between cartridges or chambers within the cartridges was observed.

Sample Stability

Verification that NPS specimens in Universal Transport Medium (UTM) at the specified conditions does not impact the performance of SARS-CoV-2 when tested with QIAstat-Dx Respiratory Panel Plus compared to freshly tested samples was conducted. A Positive sample (Table 5.8) was prepared in clinical negative matrix spiked with SARS-CoV-2 at 1x and 3x LoD concentration. Negative samples consisted of a clinical negative matrix that were prepared by pooling human negative NPS samples.

Straininformation	Source	Catalogue ID	Stock concentration
n/a	Vall d'Hebron hospital –Clinical sample	S1231	1.90E+07 copies/mL

Table 5 8. Samples used for Sample Stability Study

| Clinical sample

Sample stability testing demonstrated that NPS specimens in Universal Transport Medium (UTM) may be stored at the conditions listed below.

Room temperature up to 4 hours at 15-25°C
Refrigerated up to 3 days at approximately 4°C
Frozen up to 14 days at –20℃ ●

Matrix Equivalency and Single-spiked vs. Multi-spiked Sample Equivalency

A comparison of the performance of analytical SARS-CoV-2 samples prepared in negative clinical NPS matrix to samples prepared in simulated NPS matrix was conducted. In addition, a multi-spiked sample consisting of a subset of original panel analytes was tested together with the SARS-CoV-2 analyte.

The detection limit (1x LoD) for the SARS-CoV-2 pathogen strain defined in clinical NPS matrix as well as one log below the LoD (0.1x LoD) were taken as the reference. Samples were prepared at both concentrations in negative clinical NPS matrix and simulated NPS matrix. 20 replicates were tested for each sample concentration and matrix type using one lot of OIAstat-Dx Respiratory Panel Plus cartridges.

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The multi-spiked sample was prepared in simulated matrix at the limit of detection concentration (1x LoD) and one log below (0.1x LoD) from the cleared device (OIAstat-Dx Respiratory Panel, K183597) of each target spiked, along with the SARS-CoV-2 analyte. Twenty replicates were tested for each sample concentration using one lot of QIAstat-Dx Respiratory Panel Plus cartridges.

The matrix equivalency for the SARS-CoV-2 target was confirmed between clinical and simulated NPS matrix. Furthermore, it was confirmed that the performance was not affected when testing the additional analyte (SARS-CoV-2) in a multi-spiked with a subset of original panel analytes.

Precision

Within-laboratory precision was assessed during a study which examined the betweeninstrument variability at one site by varying test conditions such as cartridge lot, operator and instrument, since the between-site reproducibility has been tested previously with the cleared device (QIAstat-Dx Respiratory Panel, K183597). In addition, only the SARS-CoV-2 target and a subset of original panel analytes were included in the study to bridge the two devices. The testing was performed considering the following variables: across 5 non-consecutive days; two operators; three instruments; total of 90 replicates per sample concentration; sample concentrations of 3x LoD, 1x LoD, and Negative; and three cartridge lots. Analytical combined samples composed of SARS-CoV-2 and a subset of analytes were made by spiking the targets in simulated NPS matrix consisting of UTM and HeLa cells.

Table 5.9 summarizes the results, which met the acceptance criteria.

Grouping Variable(s)		Proportion		Two-Sided 95% ConfidenceLimit
Pathogen	Concentration	Fraction	Percentage	Lower	Upper
Flu B	1xLoD	89 / 90	98.89%	93.96%	99.97%
	3xLoD	91 / 92*	98.91%	94.09%	99.97%
	Neg	90 / 90	100.00%	95.98%	100.00%
CorHKU1	1xLoD	90 / 90	100.00%	95.98%	100.00%
	3xLoD	92 / 92*	100.00%	96.07%	100.00%
	Neg	90 / 90	100.00%	95.98%	100.00%
PIV 3	1xLoD	88 / 90	97.78%	92.20%	99.73%
	3xLoD	92 / 92*	100.00%	96.07%	100.00%
	Neg	90 / 90	100.00%	95.98%	100.00%
Rhinovirus	1xLoD	90 / 90	100.00%	95.98%	100.00%
	3xLoD	92 / 92*	100.00%	96.07%	100.00%
	Neg	90 / 90	100.00%	95.98%	100.00%
Adenovirus	1xLoD	87 / 90	96.67%	90.57%	99.31%

Table 5.9: Detection rate per analyte and the 2-sided 95% Confidence Interval by target for 1x LoD, 3x LoD and Negative samples

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Grouping Variable(s)		Proportion		Two-Sided 95% ConfidenceLimit
Pathogen	Concentration	Fraction	Percentage	Lower	Upper
	3xLoD	92 / 92*	100.00%	96.07%	100.00%
	Neg	90 / 90	100.00%	95.98%	100.00%
M.pneumoniae	1xLoD	86 / 90	95.56%	89.01%	98.78%
	3xLoD	90 / 92*	97.83%	92.37%	99.74%
	Neg	90 / 90	100.00%	95.98%	100.00%
SARS-CoV-2	1xLoD	87 / 90	96.67%	90.57%	99.31%
	3xLoD	92 / 92*	100.00%	96.07%	100.00%
	Neg	90 / 90	100.00%	95.98%	100.00%

*Three cartridges showed partial inhibition. Two additional cartridges were run per affected pathogen.

Conclusions

The submitted information in this premarket notification is complete and supports a substantial equivalence determination to the predicate device.

Regulation Number and Section

§ 866.3981 Device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test.

(a)
Identification. A device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test is an in vitro diagnostic device intended for the detection and identification of SARS-CoV-2 and other microbial agents when in a multi-target test in human clinical respiratory specimens from patients suspected of respiratory infection who are at risk for exposure or who may have been exposed to these agents. The device is intended to aid in the diagnosis of respiratory infection in conjunction with other clinical, epidemiologic, and laboratory data or other risk factors.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The intended use in the labeling required under § 809.10 of this chapter must include a description of the following: Analytes and targets the device detects and identifies, the specimen types tested, the results provided to the user, the clinical indications for which the test is to be used, the specific intended population(s), the intended use locations including testing location(s) where the device is to be used (if applicable), and other conditions of use as appropriate.
(2) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt (standalone or as part of a test system) for the collection of specimen types claimed by this device; alternatively, the sample collection device must be cleared in a premarket submission as a part of this device.
(3) The labeling required under § 809.10(b) of this chapter must include:
(i) A detailed device description, including reagents, instruments, ancillary materials, all control elements, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens;
(ii) Detailed descriptions of the performance characteristics of the device for each specimen type claimed in the intended use based on analytical studies including the following, as applicable: Limit of Detection, inclusivity, cross-reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, precision, reproducibility, and clinical studies;
(iii) Detailed descriptions of the test procedure(s), the interpretation of test results for clinical specimens, and acceptance criteria for any quality control testing;
(iv) A warning statement that viral culture should not be attempted in cases of positive results for SARS-CoV-2 and/or any similar microbial agents unless a facility with an appropriate level of laboratory biosafety (
e.g., BSL 3 and BSL 3+, etc.) is available to receive and culture specimens; and(v) A prominent statement that device performance has not been established for specimens collected from individuals not identified in the intended use population (
e.g., when applicable, that device performance has not been established in individuals without signs or symptoms of respiratory infection).(vi) Limiting statements that indicate that:
(A) A negative test result does not preclude the possibility of infection;
(B) The test results should be interpreted in conjunction with other clinical and laboratory data available to the clinician;
(C) There is a risk of incorrect results due to the presence of nucleic acid sequence variants in the targeted pathogens;
(D) That positive and negative predictive values are highly dependent on prevalence;
(E) Accurate results are dependent on adequate specimen collection, transport, storage, and processing. Failure to observe proper procedures in any one of these steps can lead to incorrect results; and
(F) When applicable (
e.g., recommended by the Centers for Disease Control and Prevention, by current well-accepted clinical guidelines, or by published peer-reviewed literature), that the clinical performance may be affected by testing a specific clinical subpopulation or for a specific claimed specimen type.(4) Design verification and validation must include:
(i) Detailed documentation, including performance results, from a clinical study that includes prospective (sequential) samples for each claimed specimen type and, as appropriate, additional characterized clinical samples. The clinical study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained using a comparator that FDA has determined is appropriate. Detailed documentation must include the clinical study protocol (including a predefined statistical analysis plan), study report, testing results, and results of all statistical analyses.
(ii) Risk analysis and documentation demonstrating how risk control measures are implemented to address device system hazards, such as Failure Modes Effects Analysis and/or Hazard Analysis. This documentation must include a detailed description of a protocol (including all procedures and methods) for the continuous monitoring, identification, and handling of genetic mutations and/or novel respiratory pathogen isolates or strains (
e.g., regular review of published literature and periodic in silico analysis of target sequences to detect possible mismatches). All results of this protocol, including any findings, must be documented and must include any additional data analysis that is requested by FDA in response to any performance concerns identified under this section or identified by FDA during routine evaluation. Additionally, if requested by FDA, these evaluations must be submitted to FDA for FDA review within 48 hours of the request. Results that are reasonably interpreted to support the conclusion that novel respiratory pathogen strains or isolates impact the stated expected performance of the device must be sent to FDA immediately.(iii) A detailed description of the identity, phylogenetic relationship, and other recognized characterization of the respiratory pathogen(s) that the device is designed to detect. In addition, detailed documentation describing how to interpret the device results and other measures that might be needed for a laboratory diagnosis of respiratory infection.
(iv) A detailed device description, including device components, ancillary reagents required but not provided, and a detailed explanation of the methodology, including molecular target(s) for each analyte, design of target detection reagents, rationale for target selection, limiting factors of the device (
e.g., saturation level of hybridization and maximum amplification and detection cycle number, etc.), internal and external controls, and computational path from collected raw data to reported result (e.g., how collected raw signals are converted into a reported signal and result), as applicable.(v) A detailed description of device software, including software applications and hardware-based devices that incorporate software. The detailed description must include documentation of verification, validation, and hazard analysis and risk assessment activities, including an assessment of the impact of threats and vulnerabilities on device functionality and end users/patients as part of cybersecurity review.
(vi) For devices intended for the detection and identification of microbial agents for which an FDA recommended reference panel is available, design verification and validation must include the performance results of an analytical study testing the FDA recommended reference panel of characterized samples. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(vii) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens, the design verification and validation must include a detailed description of the identity, phylogenetic relationship, or other recognized characterization of the Influenza A and B viruses that the device is designed to detect, a description of how the device results might be used in a diagnostic algorithm and other measures that might be needed for a laboratory identification of Influenza A or B virus and of specific Influenza A virus subtypes, and a description of the clinical and epidemiological parameters that are relevant to a patient case diagnosis of Influenza A or B and of specific Influenza A virus subtypes. An evaluation of the device compared to a currently appropriate and FDA accepted comparator method. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(5) When applicable, performance results of the analytical study testing the FDA recommended reference panel described in paragraph (b)(4)(vi) of this section must be included in the device's labeling under § 809.10(b) of this chapter.
(6) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens in addition to detection of SARS-CoV-2 and similar microbial agents, the required labeling under § 809.10(b) of this chapter must include the following:
(i) Where applicable, a limiting statement that performance characteristics for Influenza A were established when Influenza A/H3 and A/H1-2009 (or other pertinent Influenza A subtypes) were the predominant Influenza A viruses in circulation.
(ii) Where applicable, a warning statement that reads 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.
(iii) Where the device results interpretation involves combining the outputs of several targets to get the final results, such as a device that both detects Influenza A and differentiates all known Influenza A subtypes that are currently circulating, the device's labeling must include a clear interpretation instruction for all valid and invalid output combinations, and recommendations for any required followup actions or retesting in the case of an unusual or unexpected device result.
(iv) A limiting statement that if a specimen yields a positive result for Influenza A, but produces negative test results for all specific influenza A subtypes intended to be differentiated (
i.e., H1-2009 and H3), this result requires notification of appropriate local, State, or Federal public health authorities to determine necessary measures for verification and to further determine whether the specimen represents a novel strain of Influenza A.(7) If one of the actions listed at section 564(b)(1)(A) through (D) of the Federal Food, Drug, and Cosmetic Act occurs with respect to an influenza viral strain, or if the Secretary of Health and Human Services determines, under section 319(a) of the Public Health Service Act, that a disease or disorder presents a public health emergency, or that a public health emergency otherwise exists, with respect to an influenza viral strain:
(i) Within 30 days from the date that FDA notifies manufacturers that characterized viral samples are available for test evaluation, the manufacturer must have testing performed on the device with those influenza viral samples in accordance with a standardized protocol considered and determined by FDA to be acceptable and appropriate.
(ii) Within 60 days from the date that FDA notifies manufacturers that characterized influenza viral samples are available for test evaluation and continuing until 3 years from that date, the results of the influenza emergency analytical reactivity testing, including the detailed information for the virus tested as described in the certificate of authentication, must be included as part of the device's labeling in a tabular format, either by:
(A) Placing the results directly in the device's labeling required under § 809.10(b) of this chapter that accompanies the device in a separate section of the labeling where analytical reactivity testing data can be found, but separate from the annual analytical reactivity testing results; or
(B) In a section of the device's label or in other labeling that accompanies the device, prominently providing a hyperlink to the manufacturer's public website where the analytical reactivity testing data can be found. The manufacturer's website, as well as the primary part of the manufacturer's website that discusses the device, must provide a prominently placed hyperlink to the website containing this information and must allow unrestricted viewing access.