QUIDEL MOLECULAR INFLUENZA A + B ASSAY

K112172 · Quidel Corp. · OCC · Dec 22, 2011 · Microbiology

Device Facts

• Record ID: K112172
• Device Name: QUIDEL MOLECULAR INFLUENZA A + B ASSAY
• Applicant: Quidel Corp.
• Product Code: OCC · Microbiology
• Decision Date: Dec 22, 2011
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3980
• Device Class: Class 2

Intended Use

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. The 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 2010-2011 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 Story

Multiplex RT-PCR assay; detects influenza A (matrix protein gene) and influenza B (neuraminidase gene) viral RNA. Input: nasal/nasopharyngeal swabs; RNA extracted via bioMérieux NucliSENS easyMAG. Amplification/detection performed on Applied Biosystems 7500 Fast Dx instrument. Uses TaqMan chemistry; enzyme with reverse transcriptase, DNA polymerase, and 5'-3' exonuclease activity. Output: qualitative detection/differentiation of influenza A and B. Used in clinical laboratory settings by trained personnel. Results aid differential diagnosis alongside clinical/epidemiological factors. Benefits: rapid, sensitive molecular identification of influenza viruses to guide patient management.

Clinical Evidence

Prospective study (N=668) and retrospective study (N=372) compared subject device to FDA-cleared molecular comparator. Prospective Influenza A: 100% PPA, 98.5% NPA. Prospective Influenza B: 95.5% PPA, 97.8% NPA. Retrospective Influenza A: 100% PPA, 100% NPA. Retrospective Influenza B: 97.4% PPA, 99.4% NPA. Discordant analysis performed via sequencing.

Technological Characteristics

Multiplex Real-Time RT-PCR; TaqMan chemistry. Targets: Influenza A matrix gene, Influenza B neuraminidase gene. Instrumentation: Applied Biosystems 7500 Fast Dx (software v1.4). Extraction: bioMérieux NucliSENS easyMAG (software v2.0). Reagents: lyophilized Master Mix (primers, probes, dNTPs, enzymes). Detection: FAM (Flu A), CAL Fluor Orange 560 (Flu B), Quasar 670 (Process Control).

Indications for Use

Indicated for symptomatic patients requiring differential diagnosis of influenza A and B viral infections via nasal or nasopharyngeal swabs. Not for detection of influenza C.

Regulatory Classification

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.

Special Controls

*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.

Predicate Devices

• Prodesse ProFlu+ (k073029, k092500, k081030)

Submission Summary (Full Text)

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DEC 2 2011

# Quidel Corporation

#### Quidel Molecular Influenza A + B Assay

10/26/11 Page 1 of 16

# 510(k) Summary

#### Applicant:

Quidel Corporation 10165 McKellar Court San Diego, California 92121 Telephone: [phone] Fax: [phone]

#### 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 [phone] - Fax [e-mail]

#### Date of preparation of 510(k) summary:

December 20, 2011

### Device Name:

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

#### Legally marketed devices to which equivalence is claimed:

#### Gen-Probe Prodesse ProFlu+ (k092500)

The ProFluTM+ 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:

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 2010-2011 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:

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The Quidel Molecular Influenza A+B Assay detects viral nucleic acids that have been extracted from a patient sample using the NucliSENS® easyMAG® automated extraction platform. A multiplex real-time 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 Applied Biosystems® 7500 Fast Dx platform. Identification of influenza A occurs by the use of target specific primers and a fluorescentlabeled 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 fluorescent-labeled 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 swab and nasopharyngeal swab specimens using standard techniques from symptomatic patients. These specimens are transported, stored, and processed according to established laboratory procedures.
- 2. Nucleic Acid Extraction: Extract Nucleic Acids from the specimens with the NucliSENS easyMAG System following the manufacturer's instructions using the appropriate reagents. Use of other extraction systems with the Quidel Molecular Influenza A+B kit has not been validated. Validation of these systems is the responsibility of the end-user. Prior to the extraction procedure add 20 uL 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.
- 3. 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.
- 4. Nucleic Acid Amplification and Detection: Add 15 uL of the rehydrated Master Mix to each reaction plate well. 5uL of extracted nucleic acids (specimen with PRC) is then added to the plate well. Then place the plate into the ABI 7500 FastDx.

Once the plate is added to the instrument, the assay protocol is initiated. This protocol initiates reverse transcription of the RNA targets generating

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complementary DNA, and the subsequent amplification of the target amplicons occur. 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 35 cycles during the data collection stage of amplification, 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 the table below:

| Device Comparison | | |
|-------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Item | Subject Device<br>Quidel Molecular Influenza<br>A+B Assay | Comparator Device<br>Prodesse ProFlu+ |
| Intended Use | The Quidel Molecular<br>Influenza A+B assay is a<br>multiplex Real Time RT-PCR<br>assay for the in vitro<br>qualitative detection and<br>differentiation of influenza A<br>and influenza B viral RNA in<br>nasal and nasopharyngeal<br>swabs from patients with signs<br>and symptoms of respiratory<br>infection. This test is intended<br>for use as an aid in the<br>differential diagnosis of<br>influenza A and influenza B<br>viral infections in humans in<br>conjunction with clinical and<br>epidemiological risk factors.<br>The assay does not detect the<br>presence of influenza C virus. | The ProFlu™+ Assay is a<br>multiplex Real-Time PCR<br>(RT-PCR) in vitro diagnostic<br>test for the rapid and<br>qualitative detection and<br>discrimination of Influenza A<br>Virus, Influenza B Virus, and<br>Respiratory Syncytial Virus<br>(RSV) nucleic acids isolated<br>and purified from<br>nasopharyngeal (NP) swab<br>specimens obtained from<br>symptomatic patients. This<br>test is intended for use to aid<br>in the differential diagnosis of<br>Influenza A, Influenza B and<br>RSV viral infections in<br>humans and is not intended to<br>detect Influenza C. |
| Device Comparison | | |
| Item | Subject Device<br>Quidel Molecular Influenza<br>A+B Assay | Comparator Device<br>Prodesse ProFlu+ |
| | Negative results do not<br>preclude Influenza virus<br>infection and should not be<br>used as the sole basis for<br>diagnosis, treatment or other<br>patient management decisions.<br>Performance characteristics for<br>influenza A were established<br>during the 2010-2011<br>influenza season when | Negative results do not<br>preclude influenza or RSV<br>virus infection and should no<br>be used as the sole basis for<br>treatment or other<br>management decisions. It is<br>recommended that negative<br>RSV results be confirmed by<br>culture.<br>Performance characteristics |
| | influenza A/H3 and 2009<br>H1N1 influenza were the<br>predominant influenza A<br>viruses in circulation. When<br>other influenza A viruses are<br>emerging, performance<br>characteristics may vary. | for Influenza A Virus were<br>established when Influenza<br>A/H3 and A/H1 were the<br>predominant Influenza A<br>viruses in circulation. When<br>other Influenza A viruses are<br>emerging, performance<br>characteristics may vary. |
| | If infection with a novel<br>influenza A virus is suspected<br>based on current clinical and<br>epidemiological screening<br>criteria recommended by<br>public health authorities,<br>specimens should be collected<br>with appropriate infection<br>control precautions for novel<br>virulent Influenza viruses and<br>sent to state or local health<br>department for testing. Viral<br>culture should not be<br>attempted in these cases unless<br>a BSL 3+ facility is available<br>to receive and culture | If infection with a novel<br>Influenza A virus is suspected<br>based on current clinical and<br>epidemiological screening<br>criteria recommended by<br>public health authorities,<br>specimens should be collected<br>with appropriate infection<br>control precautions for novel<br>virulent Influenza viruses and<br>sent to state or local health<br>department for testing. Viral<br>culture should not be<br>attempted in these cases<br>unless a BSL 3+ facility is<br>available to receive and |
| Device Comparison | | |
| Item | Subject Device<br>Quidel Molecular Influenza<br>A+B Assay | Comparator Device<br>Prodesse ProFlu+ |
| Assay Target | Influenza A virus, influenza B<br>virus | Influenza A virus, influenza B<br>virus; respiratory syncytial<br>virus |
| Sample Types | nasal swab and<br>nasopharyngeal swab | nasopharyngeal swab |
| Extraction<br>Methods | bioMérieux easyMAG<br>Automated Magnetic<br>Extraction Reagents | Roche MagNA Pure LC Total<br>Nucleic Acid Isolation Kit or<br>the bioMérieux easyMAG<br>Automated Magnetic<br>Extraction Reagents |
| Assay<br>Methodology | PCR-based system for<br>detecting the presence or<br>absence of viral RNA in<br>clinical specimens | PCR-based system for<br>detecting the presence or<br>absence of viral RNA in<br>clinical specimens |
| Detection<br>Techniques | Multiplex assay using different<br>reporter dyes for each target | Multiplex assay using<br>different reporter dyes for<br>each target |
| Viral Targets | Influenza A: Matrix Gene;<br>Influenza B: conserved<br>influenza B sequence within<br>the neuraminidase gene | Influenza A: Matrix Gene;<br>Influenza B: Non-structural<br>NS1 and NS2 |
| LoD | The analytical sensitivity (limit<br>of detection or LoD) of the<br>Quidel Molecular Influenza<br>A+B assay was determined<br>using quantified (TCID50/mL)<br>cultures of 3 influenza A<br>strains (1 H1N1, 1 2009H1N1<br>and 1 H3N2), 3 influenza B<br>strains, serially diluted in<br>negative nasopharyngeal<br>matrix. Each dilution was<br>extracted using the NucliSENS<br>easyMAG System and tested<br>in replicates of 20 per | The analytical sensitivity<br>(limit of detection or LoD) of<br>the ProFlu+ Assay was<br>determined using quantified<br>(TCID50/mL) cultures of 4<br>Influenza A (2 H1N1 and 2<br>H3N2), 2 Influenza B, 2<br>Respiratory Syncytial Virus<br>Type A, and 2 Respiratory<br>Syncytial Virus Type B<br>strains serially diluted in<br>nasopharyngeal clinical<br>matrix. Each viral strain was<br>extracted using the Roche |
| Item | Subject Device<br>Quidel Molecular Influenza<br>A+B Assay | Comparator Device<br>Prodesse ProFlu+ |
| | concentration of virus on the<br>Applied Biosystems® 7500<br>Fast Dx platform. | MagNA Pure LC instrument<br>and tested in replicates of 20<br>per concentration of virus. |
| | Analytical sensitivity (LoD),<br>as defined as the lowest<br>concentration at which 95% of<br>all replicates tested positive,<br>ranged from 10¹ to 10⁰<br>TCID₅₀/mL. | Analytical sensitivity (LoD),<br>as defined as the lowest<br>concentration at which 95%<br>of all replicates tested<br>positive, ranged from 10² to<br>10⁻¹ TCID₅₀/mL. |

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Quidel Molecular Influenza A + B Assay

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# 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 included medium (5x LoD), low (2x LoD), high negative (.3x LoD) levels of influenza A virus, influenza B virus, 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 panels and controls were extracted using the bioMérieux easyMAG system and tested on the ABI 7500Fast Dx.

| Reproducibility Results | | | | | | | | | | |
|-----------------------------------------------|---------|-----------------------------------|-----|---------------------------------|-----------|-----|---------|-----------|-----|------------------|
| Panel<br>Member ID<br>(TCID50/mL) | Site 1 | | | Site 2 | | | Site 3 | | | Total<br>Results |
| | Results | AVE<br>Ct | %CV | Results | AVE<br>Ct | %CV | Results | AVE<br>Ct | %CV | |
| Influenza A<br>High<br>Negative<br>(1.44E+01) | 4/30 | 34.03*<br>(4 positive<br>results) | 2.0 | 8/30<br>(8 positive<br>results) | 34.07 | 1.9 | 0/30 | N/A | N/A | 12/90 |
| Influenza A<br>Low Positive<br>(9.6E+01) | 30/30 | 27.3 | 3.5 | 30/30 | 27.3 | 6.3 | 30/30 | 29.2 | 7.0 | 90/90 |
| Influenza A<br>Med Positive<br>(2.4E+02) | 30/30 | 25.3 | 2.9 | 30/30 | 25.2 | 5.1 | 30/30 | 26.8 | 5.5 | 90/90 |
| Influenza A<br>Negative | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 |

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| Reproducibility Results | Site 1 | | | Site 2 | | | Site 3 | | | Total Results |
|----------------------------------------------|---------|--------|-----|---------------------------------|--------|-----|---------|--------|-----|---------------|
| Panel<br>Member ID<br>(TCID50/mL) | Results | AVE Ct | %CV | Results | AVE Ct | %CV | Results | AVE Ct | %CV | |
| Influenza B<br>High<br>Negative<br>(1.3E+01) | 0/30 | N/A | N/A | 3/30<br>(3 positive<br>results) | 34.2 | 1.2 | 0/30 | N/A | N/A | 3/90 |
| Influenza B<br>Low Positive<br>(8.6E+01) | 30/30 | 24.7 | 2.6 | 30/30 | 24.6 | 4.8 | 30/30 | 25.7 | 5.1 | 90/90 |
| Influenza B<br>Med Positive<br>(2.2E+02) | 30/30 | 22.9 | 2.0 | 30/30 | 22.7 | 2.6 | 30/30 | 23.5 | 2.9 | 90/90 |
| Influenza B<br>Negative | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 |
| Influenza A<br>Positive<br>Control | 30/30 | 12.4 | 1.6 | 30/30 | 11.8 | 2.2 | 30/30 | 12.1 | 1.1 | 90/90 |
| Influenza B<br>Positive<br>Control | 30/30 | 15.2 | 2.6 | 30/30 | 14.9 | 3.1 | 30/30 | 15.1 | 1.4 | 90/90 |
| Negative<br>Control | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 |

* Ct values for positive tests

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 ABI 7500 Fast Dx.

## Limit of Detection

The analytical sensitivity (limit of detection or LoD) of the Quidel Molecular Influenza A+B assay was determined using quantified (TCID50/mL) cultures of 3 influenza A strains (1 HIN1, 1 2009H1N1 and 1 H3N2), and 3 influenza B strains, serially diluted in negative nasopharyngeal matrix. Each dilution was extracted in replicates of 20 per concentration of virus using the NucliSENS easyMAG System and tested on the Applied Biosystems® 7500 Fast Dx platform. Analytical sensitivity (LoD) is defined as the lowest concentration at which 95% of all replicates tested positive.

| Final TCID50/mL LoD | |
|---------------------|----------|
| Strain | 7500 Dx |
| A1/Mal/302/54 | 1.60E+01 |
| A/Mexico/4108/2009 | 4.80E+01 |
| A/Victoria/3/75 | 9.20E+01 |
| B/Florida/04/2006 | 4.30E+01 |
| B/RCHIN 8/05 | 1.20E+01 |
| B/Malaysia/25/06/04 | 5.70E+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 Quidel 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.

| Clinical Panel Influenza A viruses | | | | |
|------------------------------------|------------------------------|-----------|-----------|----------|
| | | | (7500 Dx) | |
| Subtype | Strain | TCID50/mL | A | B |
| H1N1 | H1N1<br>A/California/07/2009 | 1.45E+02 | Positive | Negative |
| H1N1 | A/New<br>Caledonia/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 |
| 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/Solomon<br>Islands/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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| Clinical Panel Influenza A viruses | | | | |
|------------------------------------|----------------------|-----------|-----------|----------|
| Subtype | Strain | TCID50/mL | (7500 Dx) | |
| | | | 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 |

| Clinical Panel Influenza B viruses | | | | |
|------------------------------------|-----------|-----------|----------|--|
| | | (7500 Dx) | | |
| Strain | TCID50/mL | 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 | |

| Non-clinical Restricted viruses | | | | |
|---------------------------------|--------|-----------|-----------|---|
| Subtype | Strain | TCID50/mL | (7500 Dx) | |
| | | | A | B |

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| | A/WI/629-9/2008 | 2.00E+02 | Positive | Negative |
|-------|---------------------------------------------|----------|----------|----------|
| H3N2 | A/WI/629-2/2008<br>(H3N2) | 2.00E+02 | Positive | Negative |
| H1N1 | A/WI/629-<br>S7(D02473)/2009<br>(H1N1pdm) | 2.00E+02 | Positive | Negative |
| H1N1 | A/WI/629-S5<br>(D02312)/2009<br>(H1N1pdm) | 2.00E+02 | Positive | Negative |
| H2N2 | A/Mallard/NY/6750/7<br>8 (H2N2) | 2.00E+02 | Positive | Negative |
| H7N3 | A/Chicken/NJ/15086-<br>3/94 (H7N3) | 2.00E+02 | Positive | Negative |
| H9N2 | A/Chicken/NJ/12220/<br>97 (H9N2) | 2.00E+02 | Positive | Negative |
| H4N8 | A/Mallard/OH/338/86<br>(H4N8) | 2.00E+02 | Positive | Negative |
| H6N2 | A/Chicken/CA/431/00<br>(H6N2) | 2.00E+02 | Positive | Negative |
| H8N4 | A/Blue Winged<br>Teal/LA/B174/86<br>(H8N4) | 2.00E+02 | Positive | Negative |
| H5N1 | A/Anhui/01/2005(H5<br>N1)-PR8-IBCDC-<br>RG5 | 2.00E+02 | Positive | Negative |
| H10N7 | A/GWT/LA/169GW/8<br>8 (H10N7) | 2.00E+02 | Positive | Negative |
| H11N9 | A/Chicken/NJ/15906-<br>9/96 (H11N9) | 2.00E+02 | Positive | Negative |
| H12N5 | A/Duck/LA/188D/87<br>(H12N5) | 2.00E+02 | Positive | Negative |
| H13N6 | A/Gull/MD/704/77<br>(H13N6) | 2.00E+02 | Positive | Negative |
| H14N5 | A/Mallard/GurjevRus<br>sia/262/82 (H14N5) | 2.00E+02 | Positive | Negative |
| H15N9 | A/Shearwater/Australi<br>a/2576/79 (H15N9) | 2.00E+02 | Positive | Negative |
| H16N3 | A/Shorebird/DE/172/2<br>006(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 10° to 1010 CFU/mL. Viruses were tested at concentrations of 103 to 106 TCIDs0/mL. Samples were extracted using the NucliSens easyMAG instrument and tested in

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| Quidel Molecular influenza A+B assay Cross-reactivity Data | | | |
|------------------------------------------------------------|---------------------------|-----------------------|-----------------------|
| Organism ID | CFU/mL<br>or<br>TCID50/mL | Influenza A<br>Result | Influenza B<br>Result |
| hMPV A1 | 3.70E+04 | Negative | Negative |
| hMPV BI | 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 MacIntyre 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 |
| Quidel Molecular influenza A+B assay Cross-reactivity Data | | | |
| Organism ID | CFU/mL<br>or<br>TCID50/mL | Influenza A<br>Result | Influenza B<br>Result |
| Mycobacterium avium | 1.36E+10 | Negative | Negative |
| Haemophilus influenzae | 5.90E+07 | Negative | Negative |
| 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 meningitidis | 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 diphtheriae | 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 |

triplicate. Analytical specificity of the Quidel Molecular influenza A+B assay was 100%.

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8/26/11 Page 13 of 16

# Clinical Performance:

A total of 1062 specimens were evaluated in this study (686-fresh, 376-frozen). Of the fresh specimens 373 specimens were nasal swabs, and 313 were nasopharyngeal swabs. The frozen specimens were comprised of 376 nasopharyngeal swabs.

Prospective Clinical Study

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Quidel Corporation

Ouidel Molecular Influenza A + B Assay

Six hundred and eighty six (686) fresh specimens (373 nasal swabs and 313 nasopharyngeal swabs) were tested by both the subject and comparator device for influenza A and influenza B virus viral RNA. Four 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. Seventeen specimens were invalid on initial and repeat testing (as per the device's PI) on the comparator device (2.5%). Three specimens were invalid in both devices; therefore, a total 18 specimens were removed from additional analysis. The table below details the results for the remaining 668 specimens.

| Influenza A | | | |
|----------------------------------------------------|----------|------------------------------------------|-------------------|
| Fresh nasal and<br>nasopharyngeal swabs<br>(N=668) | | Comparator: FDA Cleared RT-PCR<br>device | |
| Quidel Molecular | Positive | Negative | Total |
| Positive | 139 | 8* | 147 |
| Negative | 0 | 521 | 521 |
| Total | 139 | 529 | 668 |
| | | | <b>95% CI</b> |
| Positive Percent<br>Agreement | 139/139 | 100% | 97.4% to<br>100% |
| Negative Percent<br>Agreement | 521/529 | 98.5% | 97.0% to<br>99.3% |

*Seven specimens were negative by FDA Cleared RT-PCR device but positive for influenza A by sequence analysis. One specimen was negative by FDA Cleared RT-PCR device and negative for influenza A by sequence analysis.

| Influenza B | | | |
|----------------------------------------------------|---------------------------------------|----------|-------------------|
| Fresh nasal and<br>nasopharyngeal swabs<br>(N=668) | Comparator: FDA Cleared RT-PCR device | | |
| Quidel Molecular | Positive | Negative | Total |
| Positive | 105 | 12* | 117 |
| Negative | 5 | 546 | 551 |
| Total | 110 | 558 | 668 |
| 95% CI | | | |
| Positive Percent<br>Agreement | 105/110 | 95.5% | 89.7% to<br>98.5% |
| Negative Percent<br>Agreement | 546/558 | 97.8% | 96.3% to<br>98.9% |

*Twelve specimens were negative by FDA Cleared RT-PCR device but positive 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 1.8% (12/668) using the Quidel Molecular Influenza A + B Assay. Three of these dual infections were concordant with the FDA Cleared RT-PCR device comparator assay. Five of these dual infections were discordant with the Influenza A results from the FDA Cleared RT-PCR device comparator assay. Four of these dual infections were discordant with the Influenza B results from the FDA Cleared RT-PCR device comparator assay.

# Retrospective Study

Three hundred and seventy six (376) 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 (0.5%). 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 (0.5%). The invalid specimens were removed from performance analyses. The table below details the results for the remaining 372 specimens.

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| Influenza A | | | |
|---------------------------------------|------------------------------------------|----------|------------------|
| Frozen nasopharyngeal<br>swab (N=372) | Comparator: FDA Cleared RT-PCR<br>Device | | |
| Quidel Molecular | Positive | Negative | Total |
| Positive | 37 | 0 | 37 |
| Negative | 0 | 335 | 335 |
| Total | 37 | 335 | 372 |
| | | | 95% CI |
| Positive Percent<br>Agreement | 37/37 | 100% | 90.5% to<br>100% |
| Negative Percent<br>Agreement | 335/335 | 100% | 98.9% to<br>100% |

| | Influenza B | | |
|---------------------------------------|------------------------------------------|----------|-------------------|
| Frozen nasopharyngeal<br>swab (N=372) | Comparator: FDA Cleared RT-PCR<br>Device | | |
| Quidel Molecular | Positive | Negative | Total |
| Positive | 37 | 2* | 39 |
| Negative | 1 | 332 | 333 |
| Total | 38 | 334 | 372 |
| | | | 95% CI |
| Positive Percent<br>Agreement | 37/38 | 97.4% | 86.2% to<br>99.9% |
| Negative Percent<br>Agreement | 332/334 | 99.4% | 97.9% to<br>99.9% |

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

# CONCLUSIONS

Quidel Molecular Influenza A + B Assay yielded good positive and negative percent agreement when compared to a 510(k) cleared molecular device.

Quidel Molecular Influenza A + B Assay yielded good positive and negative percent agreement for frozen nasopharyngeal swabs compared to a 510(k) cleared molecular device.

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## DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/16/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is a stylized image of an eagle or bird-like figure with flowing lines, representing the department's mission related to health and human well-being.

Food and Drug Administration

10903 New Hampshire Avenue Silver Spring, MD 20993

Quidel Corporation c/o Ronald H. Lollar Senior Director Clinical and Quality Affairs 1055 East State Street, Suite 100 Athens, Ohio 45701

DEC 2 2 2011

Re: K112172

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: OCC, OOI Dated: November 29, 2011 Received: November 30, 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 [phone]. 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 [phone] or [phone] or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

Val, artomo

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 (if known): k112172

### Device Name: Quidel Molecular Influenza A+B Assay

#### Indication for Use:

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 2010-2011 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.

Prescription Use X (Part 21 CFR 801 Subpart D)

Over-The-Counter Use AND/OR (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 Device Evaluation and Safety (OIVD)

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Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K112172