The Quidel® Molecular Influenza A+B assay is a multiplex Real Time RT-PCR assay for the in vitro qualitative detection and differentiation of influenza A and influenza B viral RNA in nasal and nasopharyngeal swabs from patients with signs and symptoms of respiratory infection. This test is intended for use as an aid in the differential diagnosis of influenza A and influenza B viral infections in humans in conjunction with clinical and epidemiological risk factors. The assay does not detect the presence of influenza C virus.

Negative results do not preclude influenza virus infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.

Performance characteristics for influenza A were established during the 2010 to 2011 influenza season when influenza A/H3 and 2009 H1N1 influenza were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary.

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 department 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

The Quidel Molecular Influenza A+B Assay detects viral RNA that have been extracted from a patient sample using the NucliSENS® easyMAG® automated extraction platform. A multiplex RT-PCR reaction is carried out under optimized conditions in a single tube generating amplicons for each of the target viruses present in the sample. This reaction is performed utilizing the Cepheid SmartCycler® II platform. Identification of influenza A occurs by the use of target specific primers and a fluorescent- labeled probe that hybridizes to a conserved influenza A sequence within the matrix protein gene. Identification of influenza B occurs by the use of target specific primers and fluorescentlabeled probes that will hybridize to a conserved influenza B sequence within the neuraminidase gene.

The following is a summary of the procedure:

Sample Collection: Obtain nasal swabs and nasopharyngeal swabs specimens using standard techniques from symptomatic patients. These specimens are transported, stored, and processed according to established laboratory procedures.
Nucleic Acid Extraction: Extract Nucleic Acids from the specimens with the NucliSENS easyMAG System following the manufacturer's instructions using the appropriate reagents.

Prior to the extraction procedure add 20 µL of the Process Control (PRC) to each 180 uL aliquot of specimen. The PRC serves to monitor inhibitors in the extracted specimen, assures that adequate amplification has taken place and that nucleic acid extraction was sufficient.

Rehydration of Master Mix: Rehydrate the lyophilized Master Mix using 135uL of Rehydration Solution. The Master Mix contains oligonucleotide primers. fluorophore and quencher-labeled probes targeting highly conserved regions of the influenza A and influenza B viruses as well as the process control sequence. The primers are complementary to highly specific and conserved regions in the genome of these viruses. The probes are dual labeled with a reporter dye attached to the 5'end and a quencher attached to the 3' end. The rehydrated Master Mix is sufficient for eight reactions.
Nucleic Acid Amplification and Detection: Add 15 µL of the rehydrated Master Mix to each reaction tube. SuL of extracted nucleic acids (specimen with PRC) is then added to the tube. Then place the tube into the Cepheid SmartCycler® II.

Once the reaction tubes are added to the instrument, the assay protocol is initiated. This protocol initiates reverse transcription of the RNA targets generating complementary DNA, and the subsequent amplification of the target amplicons occurs. The Quidel Molecular Influenza A+B assay is based on TaqMan® chemistry, and uses an enzyme with reverse transcriptase, DNA polymerase, and 5'-3' exonuclease activities. During DNA amplification, this enzyme cleaves the probe bound to the complementary DNA sequence, separating the quencher dye from the reporter dye. This step generates an increase in fluorescent signal upon excitation by a light source of the appropriate wavelength. With each cycle, additional dye molecules are separated from their quenchers resulting in additional signal. If sufficient fluorescence is achieved by 45 cycles, the sample is reported as positive for the detected nucleic acid.

AI/ML Overview

Here's a breakdown of the acceptance criteria and the study results for the Quidel Molecular Influenza A+B Assay, based on the provided text:

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria are implicitly defined by the results of the comparative clinical study against an FDA-cleared predicate device. The performance characteristics are presented as Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA).

Performance Metric	Acceptance Criteria (Implied)	Reported Device Performance (Influenza A)	Reported Device Performance (Influenza B)
Prospective Clinical Study (Nasal & Nasopharyngeal Swabs)
PPA (Positive Agreement)	High agreement with comparator	100% (157/157)	98.4% (123/125)
NPA (Negative Agreement)	High agreement with comparator	98.7% (588/596)	95.5% (600/628)
Retrospective Clinical Study (Nasopharyngeal Swabs)
PPA (Positive Agreement)	High agreement with comparator	100% (37/37)	97.4% (37/38)
NPA (Negative Agreement)	High agreement with comparator	100% (315/315)	98.4% (309/314)

Note: The document explicitly states "good positive and negative percent agreement when compared to a high performance FDA Cleared Influenza A and B molecular test" in the conclusions, which serves as the general acceptance criterion. The precise numerical thresholds for "good" are not explicitly defined but are demonstrated by the presented results.

2. Sample Size Used for the Test Set and Data Provenance

Prospective Clinical Study:
- Sample Size: 779 fresh specimens (427 nasal swabs and 352 nasopharyngeal swabs). After removing invalid specimens, 753 specimens were analyzed.
- Data Provenance: The study was conducted during the 2010-2011 respiratory virus season (January to March 2011) at thirteen sites across the United States. The specimens were collected for routine influenza testing and tested at one central location within 72 hours of collection. This is prospective data.
Retrospective Clinical Study:
- Sample Size: 356 frozen nasopharyngeal swab specimens. After removing invalid specimens, 352 specimens were analyzed.
- Data Provenance: The study used frozen specimens collected during the 2010-2011 respiratory virus season (January to March of 2011). This is retrospective data. The country of origin is not explicitly stated but implied to be the United States, similar to the prospective study, as it's part of the same submission to the FDA in the US.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of those Experts

The ground truth for the clinical studies was established using a "high performance FDA Cleared Influenza A and B molecular test" as the comparator method. Thus, it was not established by human experts in the typical sense (e.g., radiologist consensus), but rather by the performance of an already-cleared diagnostic device.

For the discordant results in the prospective study, sequence analysis was used to resolve discrepancies for influenza A (8 specimens that were negative by comparator but positive by Quidel Molecular) and influenza B (26 specimens negative by comparator but positive by Quidel Molecular, and 2 specimens negative by comparator and negative by sequence analysis). The qualifications of those performing the sequence analysis are not detailed.

4. Adjudication Method for the Test Set

For the clinical studies, results were compared directly against the predicate FDA-cleared RT-PCR device. In cases of discordance between the subject device and the comparator device, sequence analysis was performed for some discrepant specimens (specifically, those where the comparator was negative but the Quidel Molecular assay was positive for Influenza A or B). This indicates a form of adjudication where a third, more definitive method (sequencing) was used to assess the comparator's accuracy in those specific cases, rather than an expert panel reviewing the results.

5. 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

No, an MRMC comparative effectiveness study involving human readers and AI assistance was not done. This submission is for a molecular diagnostic assay (RT-PCR), not an AI-powered image analysis or diagnostic tool that would typically involve human "readers." The comparison is between two molecular tests.

6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) was done

Yes, the device was evaluated in a standalone manner. The clinical performance data (Tables 5.8, 5.9, 5.10, 5.11) represent the Quidel Molecular Influenza A+B Assay's performance (algorithm only) compared to another FDA-cleared molecular test.

7. The Type of Ground Truth Used (expert consensus, pathology, outcomes data, etc.)

The primary "ground truth" for the clinical performance evaluation was the results of a "high performance FDA Cleared Influenza A and B molecular test" (the predicate comparator device). For specific discordant results, sequence analysis was used as a more definitive method to further evaluate the initial test results.

8. The Sample Size for the Training Set

The document does not explicitly state a sample size for a "training set" in the context of machine learning or AI. This is a molecular diagnostic assay that functions through specific primer and probe binding, not a learning algorithm that requires a distinct training and test set in the AI sense.

However, the analytical performance studies (Limit of Detection, Analytical Reactivity, Analytical Specificity) involve testing various strains and concentrations, which could be considered akin to "training" or "development" data in a broader sense for optimizing assay parameters. For example:

LoD study: Replicates of 20 per concentration for 3 influenza A strains and 3 influenza B strains.
Analytical Reactivity: 38 influenza A strains and 15 influenza B strains, tested in triplicate.
Analytical Specificity: Panels of 26 viral, 24 bacterial, and 1 yeast strain, tested in triplicate.

9. How the Ground Truth for the Training Set was Established

As this is not an AI/ML device with a conventional "training set," the concept of "ground truth for the training set" as it applies to AI is not directly applicable.

Instead, for the analytical studies:

Limit of Detection (LoD): Ground truth was established by using quantified (TCID50/mL) cultures of known influenza strains serially diluted in negative matrix. The known concentration was the "truth."
Analytical Reactivity (Inclusivity): Ground truth was established by using known, well-characterized viral strains (Influenza A subtypes and Influenza B strains) at specified TCID50 levels. The knowledge of the specific viral strain and its presence was the "truth."
Analytical Specificity (Cross-reactivity): Ground truth was established by using known concentrations of specific viral, bacterial, and yeast strains. The knowledge of which organism was present (or not present, if testing for Influenza A/B) was the "truth."

Summary

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Quidel Corporation

MAR 1 5 2012 K113777

Quidel Molecular Influenza A + B Assay

12/16/11 Page 1 of 17

Section 05, 510(k) Summary

Applicant:

Quidel Corporation 10165 McKellar Court San Diego, California 92121 Telephone: 858-552-7910 Fax: 858-646-8045

Contact Information:

Ronald H. Lollar, Senior Director Clinical and Quality Affairs 1055 East State Street Suite 100 Athens, Ohio 45701 740-589-3300 - Corporate number 740-589-3373 – Desk phone 740-593-8437 — Fax lollar@dhiusa.com

Date of preparation of 510(k) summary:

December 16, 2011

Device Name:

· Trade name - Quidel Molecular Influenza A + B Assay Classification name - Respiratory viral panel multiplex nucleic acid assay Product Code - OZE Regulation - 21 CFR 866.3980

Legally marketed devices to which equivalence is claimed:

Gen-Probe Prodesse ProFlu+ (K092500)

The ProFlu™+ Assay is a multiplex Real-Time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and discrimination of Influenza A Virus, Influenza B Virus, and Respiratory Syncytial Virus (RSV) nucleic acids isolated and purified from nasopharyngeal (NP) swab specimens obtained from symptomatic patients. This test is intended for use to aid in the differential diagnosis of Influenza A, Influenza B and RSV viral infections in humans and is not intended to detect Influenza C.

Negative results do not preclude influenza or RSV virus infection and should not be used as the sole basis for treatment or other management ·

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decisions. It is recommended that negative RSV results be confirmed by culture.

Performance characteristics for Influenza A Virus were established when Influenza A/H3 and A/H1 were the predominant Influenza A viruses in circulation. When other Influenza A viruses are emerging, performance characteristics may vary. 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 department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

Intended Use:

Negative results do not preclude influenza virus infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.

Device Description:

The Quidel Molecular Influenza A+B Assay detects viral RNA that have been extracted from a patient sample using the NucliSENS® easyMAG® automated

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extraction platform. A multiplex RT-PCR reaction is carried out under optimized conditions in a single tube generating amplicons for each of the target viruses present in the sample. This reaction is performed utilizing the Cepheid SmartCycler® II platform. Identification of influenza A occurs by the use of target specific primers and a fluorescent- labeled probe that hybridizes to a conserved influenza A sequence within the matrix protein gene. Identification of influenza B occurs by the use of target specific primers and fluorescentlabeled probes that will hybridize to a conserved influenza B sequence within the neuraminidase gene.

The following is a summary of the procedure:

1. Sample Collection: Obtain nasal swabs and nasopharyngeal swabs specimens using standard techniques from symptomatic patients. These specimens are transported, stored, and processed according to established laboratory procedures.
1. Nucleic Acid Extraction: Extract Nucleic Acids from the specimens with the NucliSENS easyMAG System following the manufacturer's instructions using the appropriate reagents.

1. Rehydration of Master Mix: Rehydrate the lyophilized Master Mix using 135uL of Rehydration Solution. The Master Mix contains oligonucleotide primers. fluorophore and quencher-labeled probes targeting highly conserved regions of the influenza A and influenza B viruses as well as the process control sequence. The primers are complementary to highly specific and conserved regions in the genome of these viruses. The probes are dual labeled with a reporter dye attached to the 5'end and a quencher attached to the 3' end. The rehydrated Master Mix is sufficient for eight reactions.
1. Nucleic Acid Amplification and Detection: Add 15 µL of the rehydrated Master Mix to each reaction tube. SuL of extracted nucleic acids (specimen with PRC) is then added to the tube. Then place the tube into the Cepheid SmartCycler® II.

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Quidel Molecular Influenza A + B Assay

based on TaqMan® chemistry, and uses an enzyme with reverse transcriptase, DNA polymerase, and 5'-3' exonuclease activities. During DNA amplification, this enzyme cleaves the probe bound to the complementary DNA sequence, separating the quencher dye from the reporter dye. This step generates an increase in fluorescent signal upon excitation by a light source of the appropriate wavelength. With each cycle, additional dye molecules are separated from their quenchers resulting in additional signal. If sufficient fluorescence is achieved by 45 cycles, the sample is reported as positive for the detected nucleic acid.

Device Comparison

The Quidel Molecular Influenza A+B assay was compared to Prodesse ProFlu+ ("Comparator Device"). The characteristics of Quidel Molecular Influenza A+B assay ("Subject Device") and Prodesse ProFlu+ ("Predicate Device") are described in Table 5.1, below:

Table 5.1: Device Comparison
Item	Subject DeviceQuidel Molecular InfluenzaA+B Assay	Comparator DeviceProdesse ProFlu+
Intended Use	The Quidel® Influenza A andB virus RT-PCR Kit is amultiplex Real Time RT-PCRassay for the in vitroqualitative detection andidentification of influenza Aand influenza B viral RNA innasal and nasopharyngealswabs from patients with signsand symptoms of respiratoryinfection. This test isintended for use as an aid inthe differential diagnosis ofinfluenza A and influenza Bviral infections in humans inconjunction with clinical andepidemiological risk factors.The assay does not detect thepresence of influenza C virus.Negative results do notpreclude influenza virus	The ProFlu™+ Assay is amultiplex Real-Time PCR(RT-PCR) in vitro diagnostictest for the rapid andqualitative detection anddiscrimination of Influenza AVirus, Influenza B Virus, andRespiratory Syncytial Virus(RSV) nucleic acids isolatedand purified fromnasopharyngeal (NP) swabspecimens obtained fromsymptomatic patients. Thistest is intended for use to aidin the differential diagnosis ofInfluenza A, Influenza B andRSV viral infections inhumans and is not intended todetect Influenza C.Negative results do notpreclude influenza or RSV
Table 5.1: Device Comparison
Item	Subject DeviceQuidel Molecular InfluenzaA+B Assay	Comparator DeviceProdesse ProFlu+
	infection and should not beused as the sole basis fordiagnosis, treatment or otherpatient management decisions.Performance characteristics forinfluenza A were establishedduring the 2010 to 2011influenza season wheninfluenza A/H3 and 2009H1N1 influenza were thepredominant influenza Aviruses in circulation. Whenother influenza A viruses areemerging, performancecharacteristics may vary.If infection with a novelinfluenza A virus is suspectedbased on current clinical andepidemiological screeningcriteria recommended bypublic health authorities,specimens should be collectedwith appropriate infectioncontrol precautions for novelvirulent influenza viruses andsent to state or local healthdepartments for testing. Viralculture should not beattempted in these cases unlessa BSL3+ facility is available toreceive and culture specimens.	virus infection and should notbe used as the sole basis fortreatment or othermanagement decisions. It isrecommended that negativeRSV results be confirmed byculture.Performance characteristicsfor Influenza A Virus wereestablished when InfluenzaA/H3 and A/H1 were thepredominant Influenza Aviruses in circulation. Whenother Influenza A viruses areemerging, performancecharacteristics may vary.If infection with a novelInfluenza A virus is suspectedbased on current clinical andepidemiological screeningcriteria recommended bypublic health authorities,specimens should be collectedwith appropriate infectioncontrol precautions for novelvirulent Influenza viruses andsent to state or local healthdepartments for testing. Viralculture should not beattempted in these casesunless a BSL 3+ facility isavailable to receive andculture specimens.
Assay Target	Influenza A virus, influenza Bvirus	Influenza A virus, influenza Bvirus, respiratory syncytialvirus
Table 5.1: Device Comparison
Item	Subject DeviceQuidel Molecular InfluenzaA+B Assay	Comparator DeviceProdesse ProFlu+
Sample Types	nasal swab andnasopharyngeal swab	nasopharyngeal swab
ExtractionMethods	bioMérieux easyMAGAutomated MagneticExtraction Reagents	Roche MagNA Pure LC TotalNucleic Acid Isolation Kit orthe bioMérieux easyMAGAutomated MagneticExtraction Reagents
AssayMethodology	PCR-based system fordetecting the presence orabsence of viral RNA inclinical specimens	PCR-based system fordetecting the presence orabsence of viral RNA inclinical specimens
DetectionTechniques	Multiplex assay using differentreporter dyes for each target	Multiplex assay usingdifferent reporter dyes foreach target
Viral Targets	Influenza A: Matrix Gene;Influenza B: NeuraminidaseGene	Influenza A: Matrix Gene;Influenza B: Non-structuralNS1 and NS2
LoD	The analytical sensitivity (limitof detection or LoD) of theQuidel Molecular InfluenzaA+B assay was determinedusing quantified (TCID50/mL)cultures of 3 influenza Astrains (1 H1N1, 1 2009H1N1and 1 H3N2), 3 influenza Bstrains, serially diluted innegative nasopharyngealmatrix. Each dilution wasextracted in replicates of 20per concentration of virususing the NucliSENSeasyMAG System and testedon Cepheid SmartCycler® II.Analytical sensitivity (LoD),	The analytical sensitivity(limit of detection or LoD) ofthe ProFlu+ Assay wasdetermined using quantified(TCID50/mL) cultures of 4Influenza A (2 H1N1 and 2H3N2), 2 Influenza B, 2Respiratory Syncytial VirusType A, and 2 RespiratorySyncytial Virus Type Bstrains serially diluted innasopharyngeal clinicalmatrix. Each viral strain wasextracted using the RocheMagNA Pure LC instrumentand tested in replicates of 20per concentration of virus.Analytical sensitivity (LoD),
Table 5.1: Device Comparison
Item	Subject DeviceQuidel Molecular InfluenzaA+B Assay	Comparator DeviceProdesse ProFlu+
	concentration at which 95% ofall replicates tested positive,ranged from 101 to 100TCID50/mL.	as defined as the lowestconcentration at which 95%of all replicates testedpositive, ranged from 102 to101 TCID50/mL.

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Quidel Moiecular Influenza A + B Assay

Analytical Performance:

Precision/Reproducibility:

The reproducibility of the Quidel Molecular Influenza A and B assay was evaluated at 3 laboratory sites. Reproducibility was assessed using a panel of 4 simulated samples that include medium (5x LoD) and low (2x LoD), high negative (0.3x LoD) influenza A (A/Mexico/4108/2009), influenza B (B/Florida/04/2006) and negative samples. Panels and controls were tested at each site by 2 operators for 5 days (triplicate testing x 2 operators x 5 days x 3 sites = 90 results per level for each virus). The LoD values are based on the values obtained in the LoD study. The panels and controls were extracted using the bioMérieux easyMAG system and tested on the Cepheid SmartCycler II.

Table 5.2: Reproducibility Results
PanelMember ID	Site 1			Site 2			Site 3			TotalResults
	Results	AVECt	%CV	Results	AVECt	%CV	Results	AVECt	%CV
Influenza AHighNegative0.3x LoD	3/30(3 positiveresults)	44.27	1.2	4/30(4positiveresults)	43.45	4.5	2/30(2 positiveresults)	42.65	0.50	9/90
Influenza ALowPositive2x LoD	30/30	37.72	2.7	30/30	38.05	3.8	30/30	37.40	4.5	90/90
Influenza AMedPositive5x LoD	30/30	35.36	1.9	30/30	36.05	3.3	30/30	34.95	1.6	90/90
Influenza ANegative	0/30	N/A	N/A	0/30	N/A	N/A	0/30	N/A	N/A	0/90
Influenza BHighNegative0.3x LoD	0/30	N/A	N/A	1/30(1positiveresults)	39.9	N/A	4/30(4 positiveresults)	42.43	3.2	4/90

Sec05_QM_FluA-B Li.doc

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Quidei Molecular Influenza A + B Assay

Table 5.2: Reproducibility Results
PanelMember ID	Site 1			Site 2			Site 3			TotalResults
	Results	AVECt	%CV	Results	AVECt	%CV	Results	AVECt	%CV
				result)
Influenza BLowPositive2x LoD	30/30	35.42	1.14	30/30	36.06	3.1	30/30	34.86	3.6	90/90
Influenza BMedPositive5x LoD	30/30	33.46	1.2	30/30	33.86	1.6	30/30	33.01	1.3	90/90
Influenza BNegative	0/30	N/A	N/A	0/30	N/A	N/A	0/30	N/A	N/A	0/90
Influenza APositiveControl	30/30	29.19	1.2	30/30	29.47	1.9	30/30	29.25	1.5	90/90
Influenza BPositiveControl	30/30	27.76	1.1	30/30	28.05	2.7	30/30	27.57	1.5	90/90
NegativeControl	0/30	N/A	N/A	0/30	N/A	N/A	0/30	N/A	N/A	0/90

The data from the combined sites indicates that the Quidel Molecular assay generates reproducible results for both influenza A and influenza B viruses when tested with the Cepheid SmartCycler II.

Limit of Detection

The analytical sensitivity (limit of detection or LoD) of the Quidel Molecular Influenza A+B assay was determined using quantified (TCIDsN/mL) cultures of three influenza A strains (1 H1N1, 1 2009H1N1 and 1 H3N2) and three influenza B strains, serially diluted in negative nasopharyngeal matrix. Each dilution was extracted using the NucliSENS easyMAG System in replicates of 20 per concentration of virus and tested on the Cepheid SmartCycler II platform. Analytical sensitivity (LoD) is defined as the lowest concentration at which 95% of all replicates tested positive.

Table 5.3: Final TCID50 LoD
Strain	Final LOD (TCID50/mL)
A1/Mal/302/54	7.00E+00
A/Mexico/4108/2009	2.40E+01
A/Victoria/3/75	3.10E+01
B/Florida/04/2006	6.00E+00
B/RCHIN 8/05	1.80E+00
B/Malaysia/25/06/04	1.30E+00

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Analytical reactivity (inclusivity)

The reactivity of the Quidel Molecular Influenza A+B assay was evaluated against multiple strains of influenza A, and influenza B viruses. The clinical panel consisted of 10 Influenza A subtype H1N1, 2 Influenza A subtype 2009H1N1, 8 Influenza A subtype H3N2, 2 Influenza A subtype H5N1, 13 Influenza B. strains. An additional panel of non-clinical restricted isolates was also tested. Each panel member was extracted using the NucliSens easyMAG instrument and tested in triplicate.

The Ouidel Molecular Influenza A+B assay detected 100% of the influenza A (38/38) and influenza B strains (15/15) at 102 to 103 TCID50 levels including novel, pandemic and avian influenza A strains and recent circulating influenza B strains.

Table 5.4: Clinical Panel Influenza A viruses
Subtype	Strain	TCID50	A	B
2009 H1N1	H1N1A/California/07/2009	1.45E+02	Positive	Negative
H1N1	A/NewCaledonia/20/1999	1.12E+02	Positive	Negative
H1N1	A/New Jersey/8/76	3.80E+02	Positive	Negative
H1N1	A/PR/8/34	5.89E+02	Positive	Negative
H1N1	A/NWS/33	NA	Positive	Negative
H1N1	A/Denver/1/57	1.26E+02	Positive	Negative
H1N1	A/FM/1/47	3.80E+02	Positive	Negative
2009 H1N1	A/Mexico/4108/2009	1.40E+02	Positive	Negative
H1N1	A1/Mal/302/54	4.19E+02	Positive	Negative
H1N1	A/Taiwan/42/06	3.39E+02	Positive	Negative
H1N1	A/Brisbane/59/07	7.24E+01	Positive	Negative
H1N1	A/SolomonIslands/3/06	1.41E+01	Positive	Negative
H3N2	A/Hong Kong/8/68	1.15E+02	Positive	Negative
H3N2	A/Wisconsin/67/2005	7.24E+02	Positive	Negative
H3N2	A/Aichi/2/68	4.17E+02	Positive	Negative

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Quidel Molecular Influenza A + B Assay

12/01/11 . Page 10 of 17

Table 5.4:	Clinical Panel Influenza A viruses
Subtype	Strain	TCID50	A	B
H3N2	A/Port Chalmers/1/73	4.57E+02	Positive	Negative
H3N2	A/Perth/16/2009	9.83E+02	Positive	Negative
H3N2	A/Uruguay/7/16/2007	1.03E+02	Positive	Negative
H3N2	A/Victoria/3/75	2.19E+02	Positive	Negative
H3N2	A/Brisbane/10/07	4.17E+02	Positive	Negative

Sec05_QM_FIuA-B Li.doc

the control control control of the control of

· .

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Quidel Molecular Influenza A + B Assay

Table 5.5:	Clinical Panel Influenza B viruses
Strain	TCID50	A	B
B/HongKong/5/72	6.67E+02	Negative	Positive
B/Panama/45/90	1.02E+02	Negative	Positive
B/Florida/02/2006	3.16E+02	Negative	Positive
B/Florida/04/2006	3.80E+02	Negative	Positive
B/Florida/07/2004	1.26E+02	Negative	Positive
B/Malaysia/25/06/04	3.41E+02	Negative	Positive
B/Maryland/1/59	1.15E+02	Negative	Positive
B/Allen/45	4.17E+02	Negative	Positive
B/Taiwan/2/62	1.51E+02	Negative	Positive
B/Russia/69	2.19E+02	Negative	Positive
B/Mass/3/66	1.38E+02	Negative	Positive
B/Lee/40	1.95E+02	Negative	Positive
B/GL/1739/54	6.30E+02	Negative	Positive

Table 5.6: Non-clinical Restricted viruses
Subtype	Strain	TCID50	A	B
	A/WI/629-9/2008	2.00E+02	Positive	Negative
H3N2	A/WI/629-2/2008(H3N2)	2.00E+02	Positive	Negative
H1N1	A/WI/629-S7(D02473)/2009(H1N1pdm)	2.00E+02	Positive	Negative
H1N1	A/WI/629-S5(D02312)/2009(H1N1pdm)	2.00E+02	Positive	Negative
H2N2	A/Mallard/NY/6750/78 (H2N2)	2.00E+02	Positive	Negative
H7N3	A/Chicken/NJ/15086-3/94 (H7N3)	2.00E+02	Positive	Negative

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Table 5.6: Non-clinical Restricted viruses
Subtype	Strain	TCID50	A	B
H9N2	A/Chicken/NJ/12220/97 (H9N2)	2.00E+02	Positive	Negative
H4N8	A/Mallard/OH/338/86(H4N8)	2.00E+02	Positive	Negative
H6N2	A/Chicken/CA/431/00(H6N2)	2.00E+02	Positive	Negative
H8N4	A/Blue WingedTeal/LA/B174/86(H8N4)	2.00E+02	Positive	Negative
H5N1	A/Anhui/01/2005(H5N1)-PR8-IBCDC-RG5	2.00E+02	Positive	Negative
H10N7	A/GWT/LA/169GW/88 (H10N7)	2.00E+02	Positive	Negative
H11N9	A/Chicken/NJ/15906-9/96 (H11N9)	2.00E+02	Positive	Negative
H12N5	A/Duck/LA/188D/87(H12N5)	2.00E+02	Positive	Negative
H13N6	A/Gull/MD/704/77(H13N6)	2.00E+02	Positive	Negative
H14N5	A/Mallard/GurjevRussia/262/82 (H14N5)	2.00E+02	Positive	Negative
H15N9	A/Shearwater/Australia/2576/79 (H15N9)	2.00E+02	Positive	Negative
H16N3	A/Shorebird/DE/172/2006(H16N3)	2.00E+02	Positive	Negative

Analytical specificity (cross-reactivity)

The analytical specificity of the Quidel Molecular Influenza A+B assay was evaluated by testing a panel consisting of 26 viral, 24 bacterial, and 1 yeast strain representing common respiratory pathogens or flora commonly present in nasopharynx. Bacteria and yeast were tested at concentrations of 105 to 1010 CFU/mL. Viruses were tested at concentrations of 103 to 10 TCID50/mL. Samples were extracted using the NucliSens easyMAG instrument and tested in triplicate. Analytical specificity of the Quidel Molecular influenza A+B assay was 100%.

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Table 5.7:	Quidel Molecular influenza A+B assay Cross-reactivity Data
Organism ID	Final Conc.	Influenza A Result	Influenza B Result
hMPV Al	3.70E+04	Negative	Negative
hMPV B1	2.37E+04	Negative	Negative
RSV Long	4.40E+04	Negative	Negative
RSV Washington	1.75E+03	Negative	Negative
Adenovirus 1/Adenoid 71	5.67E+04	Negative	Negative
Coronavirus 229E	1.70E+06	Negative	Negative
Coronavirus OC43	1.67E+06	Negative	Negative
Coxsackievirus B4	2.43E+06	Negative	Negative
Coxsackievirus B5/10/2006	2.28E+06	Negative	Negative
Cytomegalovirus	8.76E+05	Negative	Negative
Echovirus 7	5.38E+08	Negative	Negative
Echovirus 9	1.50E+06	Negative	Negative
Echovirus 6	1.05E+08	Negative	Negative
Echovirus 11	1.50E+05	Negative	Negative
Enterovirus 71	2.68E+03	Negative	Negative
Enterovirus 70	1.66E+05	Negative	Negative
Epstein Barr Virus	5,000cp/mL	Negative	Negative
HSV Type 1 Mac Inytre strain	1.95E+06	Negative	Negative
HSV Type 2 G strain	3.67E+06	Negative	Negative
Rubeola	3.78E+05	Negative	Negative
Mumps virus	8.43E+04	Negative	Negative
Parainfluenza Type 1	2.50E+05	Negative	Negative
Parainfluenza Type 2	2.20E+04	Negative	Negative
Parainfluenza Type 3	9.10E+05	Negative	Negative
Parainfluenza Type 4	9.57E+06	Negative	Negative
Varicella Zoster Virus	7.50E+02	Negative	Negative
Bordetella pertussis	1.04E+07	Negative	Negative
Bordetella bronchiseptica	2.55E+07	Negative	Negative
Chlamydia trachomatis	2.10E+05	Negative	Negative
Legionella pneumophila	2.05E+08	Negative	Negative
Mycobacterium intracellulare	6.90E+08	Negative	Negative
Mycobacterium tuberculosis	6.60E+07	Negative	Negative
Mycobacterium avium	1.36E+10	Negative	Negative
Haemophilus influenzae	5.90E+07	Negative	Negative
Table 5.7:	Quidel Molecular influenza A+B assay Cross-reactivity Data
Organism ID	Final Conc.	Influenza AResult	Influenza BResult
Pseudomonas aeruginosa	5.15E+07	Negative	Negative
Proteus vulgaris	2.65E+08	Negative	Negative
Proteus mirabilis	2.75E+07	Negative	Negative
Neisseria gonorrhoeae	2.15E+07	Negative	Negative
Neisseria menigitidis	1.85E+08	Negative	Negative
Neisseria mucosa	1.85E+08	Negative	Negative
Klebsiella pneumoniae	3.30E+07	Negative	Negative
Escherichia coli	6.80E+07	Negative	Negative
Moraxella catarrhalis	5.85E+07	Negative	Negative
Corynebacterium diptheriae	6.0E+05	Negative	Negative
Lactobacillus plantarum	1.03E+08	Negative	Negative
Streptococcus pneumoniae	4.5E+07	Negative	Negative
Streptococcus pyogenes	2.05E+08	Negative	Negative
Streptococcus salivarius	2.50E+06	Negative	Negative
Staphylococcus epidermidis	2.6E+07	Negative	Negative
Staphylococcus aureus	5.15E+08	Negative	Negative
Candida albicans	1.07E+06	Negative	Negative

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Clinical Performance:

Prospective Clinical Study

Performance characteristics of the Quidel Molecular Influenza A+B assay using the Cepheid SmartCycler® II instrument were established during a prospective study during the 2010-2011 respiratory virus season (January to March 2011). Samples used for this study were fresh nasal (427) and nasopharyngeal (352) swab specimens that were collected for routine influenza testing at thirteen sites across the United States. A single specimen was collected per patient and tested within 72-hours of collection at one central location.

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A comparator method (a high performance FDA Cleared Influenza A and B molecular test) was used in the evaluation of the Quidel Molecular Influenza A+B assay.

Seven hundred and seventy-nine (779) fresh specimens (427 nasal swabs and 352 nasopharyngeal swabs) were tested by both the subject and comparator device for influenza A and influenza B virus viral RNA. Twelve of these specimens were invalid on initial testing with the subject device (1.5%). Retesting of the specimens according to the Interpretation algorithm described above also yielded invalid results. Twenty-three specimens were invalid on initial and repeat testing (as per the device's PI) on the comparator device (3.0%). Nine specimens were invalid in both devices, therefore, a total of twenty-six invalid specimens have been removed from additional analysis. The table below details the results for the remaining 753 specimens.

Table 5.8: Influenza A
Fresh nasal andnasopharyngeal swabs(N=753)	Comparator: FDA Cleared RT-PCR device
Quidel Molecular	Positive	Negative	Total
Positive	157	8*	165
Negative	0	588	588
Total	157	596	753
			95% CI
Positive Percent Agreement	157/157	100%	97.7% to 100%
Negative Percent Agreement	588/596	98.7%	97.4% to 99.4%

*Eight specimens were negative by FDA Cleared RT-PCR device but positive for influenza A by sequence analysis.

Table 5.9:Influenza B
Fresh nasal andnasopharyngeal swabs(N=753)	Comparator: FDA Cleared RT-PCR device
Quidel Molecular	Positive	Negative	Total
Positive	123	28*	151
Negative	2	600	602
Total	125	628	753
			95% CI
Positive Percent Agreement	123/125	98.4%	94.3% to 99.8%
Negative Percent Agreement	600/628	95.5%	93.6% to 97.0%

*Twenty-six specimens were negative by FDA Cleared RT-PCR device but positive for influenza B by sequence analysis. Two specimens were negative by FDA Cleared RT-PCR device but negative for influenza B by sequence analysis.

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The prospective clinical study had a dual infection rate for Influenza A and Influenza B of 2.4% (18/753) using the Quidel Molecular Influenza A + B Assay. Three of these dual infections were concordant with the FDA Cleared RT-PCR device comparator assay. Three of these dual infections were discordant with the Influenza A results from the FDA Cleared RT-PCR device comparator assay. Twelve of these dual infections were discordant with the Influenza B results from the FDA Cleared RT-PCR device comparator assay.

Retrospective Study

Performance of the Quidel Molecular Influenza A+B assay using the Cepheid SmartCycler® II instrument were also evaluated during a retrospective study of frozen specimens collected during the 2010-2011 respiratory virus season (January to March of 2011). Samples tested in this study were frozen nasopharyngeal (356) swab specimens that were collected for routine influenza testing.

For this study the comparator method was high performance FDA Cleared influenza A and B molecular device.

Three hundred fifty six (356) frozen nasopharyngeal swabs were tested by both the subject and comparator devices for influenza A and influenza B virus viral RNA. Two of these specimens were invalid on initial testing with the subject device (0.6%). Re-testing of the specimens according to the Interpretation algorithm described above also yielded invalid results. Two specimens were invalid on initial and repeat testing (as per the device's PI) on the comparator device (0.6%). The invalid specimens were removed from performance analyses. The table below details the results for the remaining 352 specimens.

Table 5.10: Influenza A
Frozen nasopharyngealswab (N=352)	Comparator: FDA Cleared RT-PCR device
Quidel Molecular	Positive	Negative	Total
Positive	37	0	37
Negative	0	315	315
Total	37	315	352
			95% CI
Positive Percent Agreement	37/37	100%	90.5% to 100%
Negative Percent Agreement	315/315	100%	98.8% to 100%

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Table 5.11: Influenza B
Frozen nasopharyngealswab (N=352)	Comparator: FDA Cleared RT-PCR device
Quidel Molecular	Positive	Negative	Total
Positive	37	5*	42
Negative	1	309	310
Total	38	314	352
			95% CI
Positive Percent Agreement	37/38	97.4%	86.2% to 99.9%
Negative Percent Agreement	309/314	98.4%	96.3% to 99.5%

*Five specimens were negative by FDA Cleared RT-PCR device but positive for influenza B by sequence analysis.

CONCLUSIONS

Quidel Molecular Influenza A + B Assay using the Cepheid SmartCycler® II instrument yielded good positive and negative percent agreement when compared to a high performance FDA Cleared Influenza A and B molecular test.

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/17/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized caduceus symbol, which is a staff with two snakes coiled around it, and the text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" arranged in a circular fashion around the symbol. The text is in all caps and appears to be in a sans-serif font.

Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993

Quidel Corp. Ronald H. Lollar Sr. Director, Clinical and Quality Affairs 10165 McKellar Ct. San Diego. CA 92121

MAR 1 5 2012

Re: K113777

Trade/Device Name: Quidel® Molecular Influenza A+B Assay Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: Class II Product Code: OZE, OOI Dated: December 16, 2011 Received: December 21, 2011

Dear Mr. Lollar:

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. 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.

If your device is classified (see above) into class II (Special Controls), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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 requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820). This letter

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Page 2 - Ronald H. Lollar

will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5450. 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 the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

Hall, aittoms

Sally A. Hojvat, M. Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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510(k) Number K113777:

Device Name: Ouidel Molecular Influenza A+B Assay

Indication for Use:

The Ouidel® Molecular Influenza A+B assay is a multiplex Real Time RT-PCR assay for the in vitro qualitative detection and differentiation of influenza A and influenza B viral RNA in nasal and nasopharyngeal swabs from patients with signs and symptoms of respiratory infection. This test is intended for use as an aid in the differential diagnosis of influenza A and influenza B viral infections in humans in conjunction with clinical and epidemiological risk factors. The assay does not detect the presence of influenza C virus.

Negative results do not preclude influenza virus infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.

Performance characteristics for influenza A were established during the 2011 influenza season when influenza A/H3 and 2009 H1N1 influenza were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary.

Prescription Use X (Part 21 CFR 801 Subpart D)

AND/OR

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

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED

Concurrence of CDRH, Office of In Vitro Diagnostic Devices Evaluation and Safety (OIVD)

Tamara Feldblum
Division Sign-Off

Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K 1/3777

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.