K Number

Device Name

QUIDEL MOLECULAR INFLUENZA A + B ASSAY

Manufacturer

QUIDEL CORP.

Date Cleared

2011-12-22

(147 days)

Product Code

OCC OOI

Regulation Number

866.3980

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

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 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.

More Information

Contact

Type

Center

Panel

Date Received

Product Code

Subsequent Product Codes

Applicant Name (Manufacturer)

Device Name

Decision

Street 1

Street 2

City

State

Country Code

ZIP

Postal Code

Class Advise Committee

SSP Indicator

Third Party Reviewed

Expedited Review

Predicate Devices

K092500

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description details a standard Real Time RT-PCR assay for detecting viral RNA, which relies on established molecular biology techniques and instrumentation, not AI/ML for analysis or interpretation.

Therapeutic

No
This device is an in vitro diagnostic (IVD) device used for the qualitative detection of influenza A and B viral RNA. It aids in diagnosis but does not provide therapy or treatment.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the device is "intended for use as an aid in the differential diagnosis of influenza A and influenza B viral infections."

SaMD (Software as a Medical Device)

The device description clearly outlines the use of physical components such as reagents, a nucleic acid extraction system (NucliSENS easyMAG), and a real-time PCR platform (Applied Biosystems 7500 Fast Dx). While software is involved in running the assay protocol and analyzing the results, the device is fundamentally a laboratory-based diagnostic kit with associated hardware.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the assay is for the "in vitro qualitative detection and differentiation of influenza A and influenza B viral RNA". The term "in vitro" is a key indicator of an IVD, meaning it is used outside of a living organism, typically in a laboratory setting.
Sample Type: The assay uses "nasal and nasopharyngeal swabs from patients", which are biological specimens collected from the human body.
Purpose: The test is intended "as an aid in the differential diagnosis of influenza A and influenza B viral infections in humans". This clearly indicates its use in diagnosing a disease or condition.
Device Description: The description details a laboratory procedure involving sample collection, nucleic acid extraction, and a real-time RT-PCR reaction, all of which are typical steps in an in vitro diagnostic test.

Therefore, based on the provided information, the Quidel Molecular Influenza A+B assay fits the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for 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.

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.

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

OCC, OOI

Device Description

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.
1. 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.
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 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 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.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

nasal and nasopharyngeal swabs

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Not Found

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

Not Found

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

Not Found

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

Analytical Performance:
Precision/Reproducibility:
Study Type: Reproducibility
Sample Size: 90 results per level for each virus (triplicate testing x 2 operators x 5 days x 3 sites)
Key results: The Quidel Molecular Influenza A and B assay was evaluated at 3 laboratory sites using a panel of 4 simulated samples (medium positive (5x LoD), low positive (2x LoD), high negative (.3x LoD) levels of influenza A virus, influenza B virus, and negative samples). 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 (LoD):
Study Type: Analytical Sensitivity
Key results: The analytical sensitivity (LoD) of the Quidel Molecular Influenza A+B assay was determined using quantified (TCID50/mL) cultures of 3 influenza A strains and 3 influenza B strains, serially diluted in negative nasopharyngeal matrix. Each dilution was extracted in replicates of 20 per concentration of virus. LoD is defined as the lowest concentration at which 95% of all replicates tested positive. LoD ranged from 5.70E+00 to 9.20E+01 TCID50/mL for the tested strains.

Analytical reactivity (inclusivity):
Study Type: Analytical Reactivity
Key results: The reactivity of the Quidel Molecular Influenza A+B assay was evaluated against multiple strains of influenza A and influenza B viruses from a clinical panel (10 H1N1, 2 2009H1N1, 8 H3N2, 2 H5N1, 13 Influenza B) and an additional panel of non-clinical restricted isolates. The assay detected 100% of the influenza A (38/38) and influenza B strains (15/15) at 10^2 to 10^3 TCID50 levels including novel, pandemic and avian influenza A strains and recent circulating influenza B strains.

Analytical specificity (cross-reactivity):
Study Type: Analytical Specificity
Key results: 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. Bacteria and yeast were tested at concentrations of 10^6 to 10^10 CFU/mL. Viruses were tested at concentrations of 10^3 to 10^6 TCID50/mL. Analytical specificity of the Quidel Molecular influenza A+B assay was 100%.

Clinical Performance:
Study Type: Prospective Clinical Study
Sample Size: 686 fresh specimens (373 nasal swabs and 313 nasopharyngeal swabs). 668 specimens were used for analysis after removing invalid results.
Key results: The Quidel Molecular Influenza A + B Assay yielded good positive and negative percent agreement when compared to a 510(k) cleared molecular device. For Influenza A, Positive Percent Agreement was 100% (139/139) and Negative Percent Agreement was 98.5% (521/529). For Influenza B, Positive Percent Agreement was 95.5% (105/110) and Negative Percent Agreement was 97.8% (546/558). The study also noted a dual infection rate of 1.8% (12/668).

Study Type: Retrospective Study
Sample Size: 376 frozen nasopharyngeal swabs, 372 specimens used for analysis after removing invalid results.
Key results: 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. For Influenza A, Positive Percent Agreement was 100% (37/37) and Negative Percent Agreement was 100% (335/335). For Influenza B, Positive Percent Agreement was 97.4% (37/38) and Negative Percent Agreement was 99.4% (332/334).

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

Prospective Clinical Study (Fresh nasal and nasopharyngeal swabs, N=668):
Influenza A:
Positive Percent Agreement: 100% (139/139) (95% CI: 97.4% to 100%)
Negative Percent Agreement: 98.5% (521/529) (95% CI: 97.0% to 99.3%)

Influenza B:
Positive Percent Agreement: 95.5% (105/110) (95% CI: 89.7% to 98.5%)
Negative Percent Agreement: 97.8% (546/558) (95% CI: 96.3% to 98.9%)

Retrospective Study (Frozen nasopharyngeal swab, N=372):
Influenza A:
Positive Percent Agreement: 100% (37/37) (95% CI: 90.5% to 100%)
Negative Percent Agreement: 100% (335/335) (95% CI: 98.9% to 100%)

Influenza B:
Positive Percent Agreement: 97.4% (37/38) (95% CI: 86.2% to 99.9%)
Negative Percent Agreement: 99.4% (332/334) (95% CI: 97.9% to 99.9%)

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

Gen-Probe Prodesse ProFlu+ (K092500)

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

Regulation Number and Section

§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.

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

Summary

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: 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 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

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 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.
1. 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.
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 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

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
Quidel Molecular Influenza
A+B Assay	Comparator Device
Prodesse ProFlu+
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.	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.
Device Comparison
Item	Subject Device
Quidel Molecular Influenza
A+B Assay	Comparator Device
Prodesse ProFlu+
	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	Negative results do not
preclude influenza or RSV
virus infection and should no
be used as the sole basis for
treatment or other
management decisions. It is
recommended that negative
RSV results be confirmed by
culture.
Performance characteristics
	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.	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	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
Device Comparison
Item	Subject Device
Quidel Molecular Influenza
A+B Assay	Comparator Device
Prodesse ProFlu+
Assay Target	Influenza A virus, influenza B
virus	Influenza A virus, influenza B
virus; respiratory syncytial
virus
Sample Types	nasal swab and
nasopharyngeal swab	nasopharyngeal swab
Extraction
Methods	bioMérieux easyMAG
Automated Magnetic
Extraction Reagents	Roche MagNA Pure LC Total
Nucleic Acid Isolation Kit or
the bioMérieux easyMAG
Automated Magnetic
Extraction Reagents
Assay
Methodology	PCR-based system for
detecting the presence or
absence of viral RNA in
clinical specimens	PCR-based system for
detecting the presence or
absence of viral RNA in
clinical specimens
Detection
Techniques	Multiplex assay using different
reporter dyes for each target	Multiplex assay using
different reporter dyes for
each target
Viral Targets	Influenza A: Matrix Gene;
Influenza B: conserved
influenza B sequence within
the neuraminidase gene	Influenza A: Matrix Gene;
Influenza B: Non-structural
NS1 and NS2
LoD	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 H1N1, 1 2009H1N1
and 1 H3N2), 3 influenza B
strains, serially diluted in
negative nasopharyngeal
matrix. Each dilution was
extracted using the NucliSENS
easyMAG System and tested
in replicates of 20 per	The analytical sensitivity
(limit of detection or LoD) of
the ProFlu+ Assay was
determined using quantified
(TCID50/mL) cultures of 4
Influenza A (2 H1N1 and 2
H3N2), 2 Influenza B, 2
Respiratory Syncytial Virus
Type A, and 2 Respiratory
Syncytial Virus Type B
strains serially diluted in
nasopharyngeal clinical
matrix. Each viral strain was
extracted using the Roche
Item	Subject Device
Quidel Molecular Influenza
A+B Assay	Comparator Device
Prodesse ProFlu+
	concentration of virus on the
Applied Biosystems® 7500
Fast Dx platform.	MagNA Pure LC instrument
and tested in replicates of 20
per concentration of virus.
	Analytical sensitivity (LoD),
as defined as the lowest
concentration at which 95% of
all replicates tested positive,
ranged from 10¹ to 10⁰
TCID₅₀/mL.	Analytical sensitivity (LoD),
as defined as the lowest
concentration at which 95%
of all replicates tested
positive, ranged from 10² to
10⁻¹ TCID₅₀/mL.

Quidel Molecular Influenza A + B Assay

8/26/11
Page 6 of 16

تي

8/26/11 Page 7 of 16

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
Member ID
(TCID50/mL)	Site 1			Site 2			Site 3			Total
Results
	Results	AVE
Ct	%CV	Results	AVE
Ct	%CV	Results	AVE
Ct	%CV
Influenza A
High
Negative
(1.44E+01)	4/30	34.03*
(4 positive
results)	2.0	8/30
(8 positive
results)	34.07	1.9	0/30	N/A	N/A	12/90
Influenza A
Low Positive
(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
Med Positive
(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
Negative	0/30	N/A	N/A	0/30	N/A	N/A	0/30	N/A	N/A	0/90

Reproducibility Results	Site 1			Site 2			Site 3			Total Results
Panel
Member ID
(TCID50/mL)	Results	AVE Ct	%CV	Results	AVE Ct	%CV	Results	AVE Ct	%CV
Influenza B
High
Negative
(1.3E+01)	0/30	N/A	N/A	3/30
(3 positive
results)	34.2	1.2	0/30	N/A	N/A	3/90
Influenza B
Low Positive
(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
Med Positive
(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
Negative	0/30	N/A	N/A	0/30	N/A	N/A	0/30	N/A	N/A	0/90
Influenza A
Positive
Control	30/30	12.4	1.6	30/30	11.8	2.2	30/30	12.1	1.1	90/90
Influenza B
Positive
Control	30/30	15.2	2.6	30/30	14.9	3.1	30/30	15.1	1.4	90/90
Negative
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

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
A/California/07/2009	1.45E+02	Positive	Negative
H1N1	A/New
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
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

8/26/11
Page 10 of 16

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

	A/WI/629-9/2008	2.00E+02	Positive
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/6750/7
8 (H2N2)	2.00E+02	Positive	Negative
H7N3	A/Chicken/NJ/15086-
3/94 (H7N3)	2.00E+02	Positive	Negative
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 Winged
Teal/LA/B174/86
(H8N4)	2.00E+02	Positive	Negative
H5N1	A/Anhui/01/2005(H5
N1)-PR8-IBCDC-
RG5	2.00E+02	Positive	Negative
H10N7	A/GWT/LA/169GW/8
8 (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/GurjevRus
sia/262/82 (H14N5)	2.00E+02	Positive	Negative
H15N9	A/Shearwater/Australi
a/2576/79 (H15N9)	2.00E+02	Positive	Negative
H16N3	A/Shorebird/DE/172/2
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

Quidel Molecular influenza A+B assay Cross-reactivity Data
Organism ID	CFU/mL
or
TCID50/mL	Influenza A
Result	Influenza B
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
or
TCID50/mL	Influenza A
Result	Influenza B
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%.

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

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
nasopharyngeal swabs
(N=668)		Comparator: FDA Cleared RT-PCR
device
Quidel Molecular	Positive	Negative	Total
Positive	139	8*	147
Negative	0	521	521
Total	139	529	668
			95% CI
Positive Percent
Agreement	139/139	100%	97.4% to
100%
Negative Percent
Agreement	521/529	98.5%	97.0% to
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
nasopharyngeal swabs
(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
Agreement	105/110	95.5%	89.7% to
98.5%
Negative Percent
Agreement	546/558	97.8%	96.3% to
98.9%

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

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.

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

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

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

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,

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

510(k) Number (if known): K112172

Device Name: Quidel Molecular Influenza A+B Assay

Indication for 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.

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