Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The document describes a molecular diagnostic assay using isothermal amplification and fluorescence detection, interpreted by "on-board method-specific algorithms." There is no mention of AI, ML, or any learning-based approach for interpreting the results. The algorithms appear to be deterministic based on fluorescence signal thresholds.

Therapeutic

No.
The device is a diagnostic test used to detect the presence of influenza A and B viral RNA, providing an aid in differential diagnosis. It does not provide therapy or treatment.

Diagnostic

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states that the Solana® Influenza A+B Assay is a "qualitative in vitro diagnostic test" intended to "aid in the differential diagnosis of influenza A and influenza B viral infections." This language directly identifies its purpose as a diagnostic tool.

SaMD (Software as a Medical Device)

No

The device description explicitly states that the assay is placed in the "Solana instrument" for amplification and detection, and that the Solana instrument measures and interprets the fluorescent signal and reports results on its display screen. This indicates the device includes dedicated hardware (the Solana instrument) in addition to the software that runs on it.

IVD (In Vitro Diagnostic)

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

The "Intended Use / Indications for Use" section explicitly states: "The Solana® Influenza A+B Assay is a qualitative in vitro diagnostic test for the detection and differentiation of influenza A and influenza B viral RNA..."

This statement clearly identifies the device as an in vitro diagnostic test.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Solana® Influenza A+B Assay is a qualitative in vitro diagnostic test for the 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 the Solana® Influenza A+B Assay were established during the spring of 2016 when influenza A/H3 and 2009 H1N1 influenza were the predominant influenza viruses in circulation. When other influenza viruses are emerging, performance characteristics may vary.

If infection with a novel influenza 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

Device Description

The Solana Influenza A+B Assay amplifies and detects viral RNA present in viral transport media containing nasopharyngeal or nasal swab specimens obtained from symptomatic patients.

The assay consists of two major steps: 1) specimen preparation, and 2) amplification and detection of target sequences specific to influenza A and/or influenza B using isothermal Reverse Transcriptase - Helicase-Dependent Amplification (RT-HDA) in the presence of target-specific fluorescence probes.

A patient nasal or nasopharyngeal swab specimen in viral transport media is transferred to a Process Buffer Tube, subjected to heat treatment at 95°C for 5 minutes and mixed. The processed sample is transferred to a Reaction Tube. The Reaction Tube contains lyophilized RT-HDA reagents, dNTPs, primers and probes. Once rehydrated with the processed sample, the Reaction Tube is placed in Solana for amplification and detection of influenza A and influenza B-specific target sequences. In Solana, the target sequences are amplified by influenza A and influenza B specific primers and detected by influenza A and influenza B specific fluorescence probes, respectively. A competitive process control (PRC) is included in the Process Buffer Tube to monitor sample processing, inhibitory substances in clinical samples, reagent failure or device failure. The PRC target is amplified by influenza B specific primers and detected by a PRC specific fluorescence probe.

The two target probes and PRC probe are labeled with a quencher on one end and a fluorophore on the other end. In addition, the two target probes and PRC probe have one or more bases that are comprised of ribonucleic acid. Upon annealing to influenza A, influenza B or PRC amplicons, the fluorescence probes are cleaved by RNaseH2 and the fluorescence signal increases due to physical separation of fluorophore from quencher. Solana measures and interprets the fluorescent signal, using on-board method-specific algorithms. Solana then reports the test results to the user on its display screen, and it can print out the results via an integrated printer.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

nasal, nasopharyngeal

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

A four sample panel consisting of three levels of a combined influenza A and influenza B contrived sample and a negative contrived sample are tested in this study. Influenza A and influenza B viruses (Influenza A/California/07/2009 and Influenza B/Brisbane/60/08, respectively) are diluted in negative nasal matrix to 2 x LOD for moderate positive, 1 x LOD for low positive and diluted to C20 to C80 for high negative / low positive nasal matrix without spiked virus is used for the negative sample. The Solana Influenza A+B assay was used according to the instructions for use.

Panels and controls were tested at each site by two operators per instrument for five days, each sample tested in three (3) replicates, for a total of 90 results per level for each virus for each instrument (2 operators x 5 days x 3 sites x 3 replicates).

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

Reproducibility (Analytical performance): The reproducibility of the Solana® Influenza A+B Assay was evaluated at three laboratory sites. Panels and controls were tested at each site by two operators per instrument for five days, each sample tested in three (3) replicates, for a total of 90 results per level for each virus for each instrument (2 operators x 5 days x 3 sites x 3 replicates).

Influenza A/California/07/2009 High Negative (1.4 x102 TCID50/mL): Overall Percent Agreement 64.4% (95% CI: 54.1 to 73.6)
Influenza A/California/07/2009 Low Positive (4.7x102 TCID50/mL): Overall Percent Agreement 100% (95% CI: 96.5 to 100)
Influenza A/California/07/2009 Moderate Positive (9.4x102 TCID50/mL): Overall Percent Agreement 100% (95% CI: 96.5 to 100)
Negative: Overall Percent Agreement 100% (95% CI: 96.5 to 100)
Influenza A Positive Control: Overall Percent Agreement 100% (95% CI: 94.2 to 100)
Influenza A Negative Control: Overall Percent Agreement 100% (95% CI: 94.2 to 100)
Influenza B/Brisbane/60/08 High Negative (2.6 x101 TCID50/mL): Overall Percent Agreement 26.7% (95% CI: 18.6 to 36.6)
Influenza B/Brisbane/60/08 Low Positive (8.5x102TCID50/mL): Overall Percent Agreement 100% (95% CI: 96.5 to 100)
Influenza B/Brisbane/60/08 Moderate Positive (1.7x102 TCID50/mL): Overall Percent Agreement 100% (95% CI: 96.5 to 100)
Negative: Overall Percent Agreement 100% (95% CI: 96.5 to 100)
Influenza B Positive Control: Overall Percent Agreement 100% (95% CI: 94.2 to 100)
Influenza B Negative Control: Overall Percent Agreement 100% (95% CI: 94.2 to 100)
The results suggest that there are no significant differences between different users and different sites on different days. Reproducibility studies are acceptable.

Clinical Study (Comparison Versus Culture with DFA and DSFA): N=1473 specimens (302 nasal, 1171 nasopharyngeal) collected in viral transport media from February to April 2016 from five sites across the United States. All specimens were transported to a central location for testing by comparator methods (culture for influenza A and B using R-Mix Too mixed cells and direct specimen DFA (DSFA), and extraction with NucliSENS® easyMAG® and testing with an FDA-cleared Influenza A+B molecular assay). Fresh (742) and frozen (731) specimens were tested with Solana Influenza A+B Assay. 15 specimens were contaminated/toxic in cell culture and 50 were invalid in Solana Assay, resulting in 1408 specimens for analysis.

Influenza A:
- Fresh (N=709): Sensitivity 98.9% (95% CI: 96.1 to 99.7), Specificity 95.4% (95% CI: 93.3 to 96.9)
- Frozen (N=699): Sensitivity 98.3% (95% CI: 95.2 to 99.4), Specificity 94.8% (95% CI: 92.6 to 96.4)
- All (N=1408): Sensitivity 98.6% (95% CI: 96.8 to 99.4), Specificity 95.1% (95% CI: 93.7 to 96.3)
- 51 discordant specimens (Solana Positive/Culture and DSFA Negative), 28 were positive by an alternate molecular assay.
- 5 discordant specimens (Solana Negative/Culture and DSFA Positive), 2 were positive by an alternate FDA cleared molecular assay.
Influenza B:
- Fresh (N=709): Sensitivity 100% (95% CI: 94.2 to 100), Specificity 99.8% (95% CI: 99.1 to 100)
- Frozen (N=699): Sensitivity 100% (95% CI: 85.7 to 100), Specificity 98.8% (95% CI: 97.7 to 99.4)
- All (N=1408): Sensitivity 100% (95% CI: 95.7 to 100), Specificity 99.3% (95% CI: 98.7 to 99.6)
- 9 discordant specimens (Solana Positive/Culture and DSFA Negative), 2 were positive by an alternate FDA cleared molecular assay.

Clinical Study (Comparison with an FDA-cleared Influenza A+B Molecular Assay): N=1473 specimens. Comparison testing performed on fresh specimens within 72-hours of collection. 31 invalid in comparator assay, 50 invalid in Solana Assay (1 invalid in both), resulting in 1393 specimens for analysis.

Influenza A:
- Fresh (N=710): PPA 96.5% (95% CI: 93.0 to 98.3), NPA 98.2% (95% CI: 98.7 to 99.1)
- Frozen (N=683): PPA 97.8% (95% CI: 94.5 to 99.2), NPA 95.2% (95% CI: 92.9 to 96.7)
- All (N=1393): PPA 97.2% (95% CI: 95.0 to 98.4), NPA 96.7% (95% CI: 95.4 to 97.7)
- 44 total discordant specimens. Of 33 discordant (Solana Positive/Comparator Negative), 9 were positive by culture/DSFA. Of 11 discordant (Solana Negative/Comparator Positive), 2 were positive by culture/DSFA.
Influenza B:
- Fresh (N=710): PPA 100% (95% CI: 93.7 to 100), NPA 99.1% (95% CI: 98.0 to 99.6)
- Frozen (N=683): PPA 100% (95% CI: 85.7 to 100), NPA 98.8% (95% CI: 97.6 to 99.4)
- All (N=1393): PPA 100% (95% CI: 95.4 to 100), NPA 98.9% (95% CI: 98.2 to 99.4)
- 14 total discordant specimens (Solana Positive/Comparator Negative), 7 were positive by culture/DSFA.

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

See "Summary of Performance Studies" section.

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.

K131728

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

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & 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 three human profiles facing to the right, stacked on top of each other.

September 27, 2016

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

Ronald H. Lollar Senior Director, Clinical and Regulatory Affairs Quidel Corporation 2005 East State Street, Suite 100 Athens, OH 45701

Re: K161814

Trade/Device Name: Solana® 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. OZE Dated: June 30, 2016 Received: July 1, 2016

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. Iisting of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's 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

1

medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled. "Misbranding by reference to premarket notification" (21CFR 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 Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Steven R. Gitterman -S

for Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

2

Indications for Use

510(k) Number (if known) K161814

Device Name Solana® Influenza A+B Assay

Indications for Use (Describe)

The Solana® Influenza A+B Assay is a qualitative in vitro diagnostic test for the 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 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 spring of 2016 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.

Type of Use (Select one or both, as applicable)

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

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3

Applicant:

Quidel Corporation 12544 High Bluff Drive, Suite 200 San Diego, California 92130 Telephone: 858-552-7910 Fax: 858-646-8045

Contact Information:

Ronald H. Lollar, Senior Director Clinical and Regulatory Affairs 2005 East State Street, Suite 100 Athens, Ohio 45701 740-589-3300 - Corporate number 740-589-3373 – Desk phone 858-552-6451—Fax Ron.Lollar@quidel.com

Date of preparation of 510(k) summary:

June 30, 2016

A. 510(k) Number:

K161814

B. Purpose for Submission:

To obtain substantial equivalence for the Solana Influenza A+B Assay when performed on the Solana

C. Measurand:

Influenza A: Matrix Gene; Influenza B: Matrix Gene

D. Type of Test:

Reverse Transcriptase - Helicase-Dependent Amplification (RT-HDA)

4

E. Applicant:

Quidel Corporation

F. Proprietary and Established Names:

Solana Influenza A+B Assay

G. Regulatory Information:

Table 1.Regulatory Information
Product Code	Classification	Regulation Section	Panel
OCC	Class II	21 CFR 866.3980 Respiratory
viral panel multiplex nucleic acid
assay	Microbiology
(83)

H. Intended Use:

1. Intended Use(s):

The Solana® Influenza A+B Assay is a qualitative in vitro diagnostic test for the 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 the Solana® Influenza A+B Assay were established during the spring of 2016 when influenza A/H3 and 2009 H1N1 influenza were the predominant influenza viruses in circulation. When other influenza viruses are emerging, performance characteristics may vary.

If infection with a novel influenza 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

5

should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

1. Indication(s) for Use:
  Same as Intended Use.

3. Special conditions for use statement(s):

For in vitro diagnostic use only
For prescription use only ●

4. Special instrument requirements:

Solana® Instrument

l. Device Description:

The Solana Influenza A+B Assay amplifies and detects viral RNA present in viral transport media containing nasopharyngeal or nasal swab specimens obtained from symptomatic patients.

The assay consists of two major steps: 1) specimen preparation, and 2) amplification and detection of target sequences specific to influenza A and/or influenza B using isothermal Reverse Transcriptase - Helicase-Dependent Amplification (RT-HDA) in the presence of target-specific fluorescence probes.

A patient nasal or nasopharyngeal swab specimen in viral transport media is transferred to a Process Buffer Tube, subjected to heat treatment at 95°C for 5 minutes and mixed. The processed sample is transferred to a Reaction Tube. The Reaction Tube contains lyophilized RT-HDA reagents, dNTPs, primers and probes. Once rehydrated with the processed sample, the Reaction Tube is placed in Solana for amplification and detection of influenza A and influenza B-specific target sequences. In Solana, the target sequences are amplified by influenza A and influenza B specific primers and detected by influenza A and influenza B specific fluorescence probes, respectively. A competitive process control (PRC) is included in the Process Buffer Tube to monitor sample processing, inhibitory substances in clinical samples, reagent failure or device failure. The PRC target is amplified by influenza B specific primers and detected by a PRC specific fluorescence probe.

The two target probes and PRC probe are labeled with a quencher on one end and a fluorophore on the other end. In addition, the two target probes and PRC probe have one or more bases that are comprised of ribonucleic acid. Upon annealing to influenza A,

6

influenza B or PRC amplicons, the fluorescence probes are cleaved by RNaseH2 and the fluorescence signal increases due to physical separation of fluorophore from quencher. Solana measures and interprets the fluorescent signal, using on-board method-specific algorithms. Solana then reports the test results to the user on its display screen, and it can print out the results via an integrated printer.

Materials Provided:

Solana® Influenza A+B Assay Kit: M300 48 Tests per kit

Table 2.Kit Components
Component	Quantity	Storage
Process Buffer	48 tubes/kit 1.55 mL	2°C to 8°C
Reaction Tubes	48 tubes/kit	2°C to 8°C

Materials required but not provided:

External controls for Influenza A and Influenza B (e.g. Quidel Molecular Influenza ● A+B Control Set, which contains positive and negative controls, serves as an external processing control)
Sterile DNAse-free filter-blocked positive displacement micropipettor tips ●
Micropipettor
Stopwatch or timer ●
Scissors or a blade
Workflow tray
. Transfer Rack
Heat block capable of 95 ± 2°C temperature ●
Thermometer
Solana instrument
Transport Media (BD/Copan UTM, Remel M4, Remel M4RT, Remel M5, Remel M6, ● or Copan eSwab)

J. Substantial Equivalence Information:

1. Predicate device name(s):
  Lyra® Influenza A+B Assay
1. Predicate 510(k) number(s): K131728

7

3. Comparison with predicate:

	Table 3. Similarities
Item	Solana® Influenza A+B Assay	Lyra® Influenza A+B Assay
(K131728)
Intended Use	The Solana® Influenza A+B Assay is a
qualitative in vitro diagnostic test for the
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 spring
2016 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 | The Lyra® Influenza A+B assay
is a multiplex Real Time RT-PCR
assay for the in vitro qualitative
detection and differentiation of
influenza A and influenza B
viral RNA in nasal and
nasopharyngeal swabs from
patients with signs and
symptoms of respiratory
infection. This test is intended
for use as an aid in the
differential diagnosis of
influenza A and influenza B
viral infections in humans in
conjunction with clinical and
epidemiological risk factors.
The assay does not detect the
presence of influenza C virus.

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

Performance characteristics for
influenza A were established
during the 2011 and 2013
influenza seasons when
influenza A/H3 and 2009 H1N1 | |
| | Table 3.Similarities | | |
| Item | Solana® Influenza A+B Assay | Lyra® Influenza A+B Assay (K131728) | |
| | 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. | 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.

8

Solana® Influenza A+B Assay 6/30/2016 Page 6 of 24

510(k) Summary

9

Solana® Influenza A+B Assay 6/30/2016 Page 7 of 24

510(k) Summary

10

K. Standard/Guidance Document Referenced (if applicable):

Guidance for Industry and FDA Staff: Statistical Guidance on Reporting Results from Studies Evaluating Diagnostic Tests (Final, 3/13/2007)

11

http://www.fda.gov/downloads/MedicalDevices/DeviceRegulationandGuidance/GuidanceD ocuments/ucm071287.pdf

Guidance for Industry and FDA Staff - Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay -

http://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/GuidanceDocuments/ ucm180307.htm

Guidance on Informed Consent for In Vitro Diagnostic Device Studies Leftover Human Specimens that are Not Individually Identifiable (April 2006) – http://www.fda.gov/cdrh/oivd/guidance/1588.pdf.

Guidance for Industry and Food and Drug Administration Staff - eCopy Program for Medical Device (December 2012)

http://www.fda.gov/downloads/MedicalDevices/DeviceRegulationandGuidance/GuidanceD ocuments/UCM313794.pdf

L. Test Principle:

The Solana Influenza A+B Assay amplifies and detects viral RNA present in viral transport media containing nasopharyngeal or nasal swab specimens obtained from symptomatic patients.

The assay consists of two major steps: 1) specimen preparation, and 2) amplification and detection of target sequences specific to influenza A and/or influenza B using isothermal Reverse Transcriptase - Helicase-Dependent Amplification (RT-HDA) in the presence of target-specific fluorescence probes.

A patient nasal or nasopharyngeal swab specimen in viral transport media is transferred to a Process Buffer Tube, subjected to heat treatment at 95°C for 5 minutes and mixed. The processed sample is transferred to a Reaction Tube. The Reaction Tube contains lyophilized RT-HDA reagents, dNTPs, primers and probes. Once rehydrated with the processed sample, the Reaction Tube is placed in Solana for amplification and detection of influenza A and influenza B-specific target sequences. In Solana, the target sequences are amplified by influenza A and influenza B specific primers and detected by influenza A and influenza B specific fluorescence probes, respectively. A competitive process control (PRC) is included in the Process Buffer Tube to monitor sample processing, inhibitory substances in clinical samples, reagent failure or device failure. The PRC target is amplified by influenza B specific primers and detected by a PRC specific fluorescence probe.

12

The two target probes and PRC probe are labeled with a quencher on one end and a fluorophore on the other end. In addition, the two target probes and PRC probe have one or more bases that are comprised of ribonucleic acid. Upon annealing to influenza A, influenza B or PRC amplicons, the fluorescence probes are cleaved by RNaseH2 and the fluorescence signal increases due to physical separation of fluorophore from quencher. Solana measures and interprets the fluorescent signal, using on-board method-specific algorithms. Solana then reports the test results to the user on its display screen, and it can print out the results via an integrated printer.

M. Performance Characteristics:

1. Analytical performance:

a. Precision/Reproducibility:

Reproducibility

The reproducibility of the Solana® Influenza A+B Assay was evaluated at three laboratory sites. A four sample panel consisting of three levels of a combined influenza A and influenza B contrived sample and a negative contrived sample are tested in this study. Influenza A and influenza B viruses (Influenza A/California/07/2009 and Influenza B/Brisbane/60/08, respectively) are diluted in negative nasal matrix to 2 x LOD for moderate positive, 1 x LOD for low positive and diluted to C20 to C80 for high negative / low positive nasal matrix without spiked virus is used for the negative sample. The Solana Influenza A+B assay was used according to the instructions for use.

Panels and controls were tested at each site by two operators per instrument for five days, each sample tested in three (3) replicates, for a total of 90 results per level for each virus for each instrument (2 operators x 5 days x 3 sites x 3 replicates).

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Solana® Influenza A+B Assay 6/30/2016 Page 11 of 24

510(k) Summary

Table 5. Reproducibility Summary
	SITE		SITE		SITE
	Site #1	Site #2	Site #3	Overall Percent Agreement	95% Confidence Interval
	#Detected positive/# tested	% Agreement with Expected Result	#Detected positive/# tested			% Agreement with Expected Result	#Detected positive/# tested	% Agreement with Expected Result
Influenza A/California/07/2009 High Negative (1.4 x102 TCID50/mL)	10/30	33.3	25/30	83.3	23/30	76.7	58/90	64.4	54.1 to 73.6
Influenza A/California/07/2009 Low Positive (4.7x102 TCID50/mL)	30/30	100	30/30	100	30/30	100	90/90	100	96.5 to 100
Influenza A/California/07/2009 Moderate Positive (9.4x102 TCID50/mL)	30/30	100	30/30	100	30/30	100	90/90	100	96.5 to 100
Negative	0/30	100	0/30	100	0/30	100	0/90	100	96.5 to 100
Influenza A Positive Control	15/15	100	15/15	100	15/15	100	45/45	100	94.2 to 100
Influenza A Negative Control	0/15	100	0/15	100	0/15	100	0/45	100	94.2 to 100
	SITE		SITE		SITE
	Site #1	Site #2	Site #3	Overall Percent Agreement With Expected Results	95% Confidence Interval
	#Detected positive/# tested	% Agreement with Expected Result	#Detected positive/# tested			% Agreement with Expected Result	#Detected positive/# tested	% Agreement with Expected Result
Influenza B/Brisbane/60/08 High Negative (2.6 x101 TCID50/mL)	9/30	30	5/30	16.7	10/30	33.3	24/90	26.7	18.6 to 36.6
Influenza B/Brisbane/60/08 Low Positive (8.5x102TCID50/mL)	30/30	100	30/30	100	30/30	100	90/90	100	96.5 to 100
Influenza B/Brisbane/60/08 Moderate Positive (1.7x102 TCID50/mL)	30/30	100	30/30	100	30/30	100	90/90	100	96.5 to 100
Negative	0/30	100	0/30	100	0/30	100	0/90	100	96.5 to 100
Table 5. Reproducibility Summary
	SITE
	Site #1	Site #2	Site #3
	#Detected
positive/#
tested	% Agreement
with
Expected
Result	#Detected
positive/#
tested	%
Agreement
with
Expected
Result	#Detected
positive/#
tested	% Agreement
with Expected
Result	Overall Percent
Agreement	95%
Confidence
Interval
Influenza B Positive
Control	15/15	100	15/15	100	15/15	100	45/45	100	94.2 to 100
Influenza B Negative
Control	0/15	100	0/15	100	0/15	100	0/45	100	94.2 to 100

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The results suggest that there are no significant differences between different users and different sites on different days. Reproducibility studies are acceptable.

b. Linearity/assay reportable range:
Not applicable - This assay is qualitative.
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
Traceability:

Not applicable. This assay is qualitative.

Specimen Stability:

Influenza A and B strains (Influenza A/California/07/2009 and Influenza B/Brisbane/60/08, respectively) were formulated in six (6) transport medium pooled negative matrix (Copan UTM, Remel M4, Remel M5, Remel M6, Remel M4RT or Copan ESwab transport media) at a final concentration of 2x LOD level.

The transport media systems containing the contrived samples were stored at 2° to 8°C up to 9 days. The samples were processed according to the instructions for use. Each transport media was tested in 3 replicates at Day 0, 24 hours, 48 hours, 72 hours, Day 7, and Day 9.

Influenza A and influenza B are stable in transport media BD UTM (1- and 3- mL), Remel M4 (3-mL), Remel M4RT (3-mL), Remel M5 (3-mL), and Remel M6 (3-mL) at 2° to 8°C for up to 9 days.

15

Influenza A and influenza B are stable in Copan eSwab transport media at 2 to 8°C for up to 48 hours.

Controls:

Controls (Quidel Molecular Influenza A+B Control Set), which contains positive and negative controls, serves as an external processing and extraction control) were run on the Solana Influenza A+B Assay each day of testing. These controls are described as follows:

a. The process control is used to monitor sample processing, to detect HDA inhibitory specimens, to confirm the integrity of assay reagents and the operation of the Solana instrument. The process control is included in the Reaction Mix tube.
b. The external positive control may be treated as a patient specimen. The control should be sampled and tested as if it were a patient specimen and processed as described above in the Assay Procedure. The external positive control is intended to monitor substantial reagent and instrument failure.
c. The external negative control may be treated as a patient specimen. The control should be sampled and tested as if it were a patient specimen and processed as described above in the Assay Procedure. The external negative control is used to detect reagent or environmental contamination (or carryover) by influenza A or B RNA or amplicon.

d. Detection limit:

The analytical sensitivity (limit of detection or LOD) of the Solana® Influenza A+B Assay was determined using quantified (TCID50/mL) cultures of three (3) influenza A strains and two (2) influenza B strains, serially diluted in negative nasopharyngeal matrix. Each dilution was run as 20 replicates in the Solana Influenza A+B assay. Analytical sensitivity (LOD) is defined as the lowest concentration at which at least 95% of all replicates tested positive. The demonstrated LOD for each strain tested is shown below:

16

Table 6.LOD Values
Virus		TCID50/mL
Influenza A	Subtype
A/Taiwan/42/06	H1N1	$7.5x10^2$
A/California/07/2009	H1N1p	$4.7x10^2$
A/Texas/50/2012	H3N2	$6.3x10^0$
Influenza B	Lineage
B/Brisbane/60/08	Victoria	$8.5x10^1$
B/Massachusetts/2/2012	Yamagata	$3.3x10^1$

e. Analytical specificity:

Cross Reactivity:

A study was performed to evaluate the cross-reactivity of the Solana® Influenza A+B Assay with forty-four (44) microorganisms (24 bacteria, 1 yeast, 19 viruses) potentially found in specimens that are collected from patients symptomatic for influenza. Each microorganism was diluted in negative nasal matrix to the desired concentration (106 or higher CFU/mL for bacteria, yeast and 105 or higher pfu/mL or TCID50/mL for viruses) and tested. The organisms and their concentrations included in the cross-reactivity study are shown in the table below.

Table 7.Cross-reactive Organisms
Organism	Concentration
Tested
Adenovirus 1	1.0x105 TCID50/mL
Adenovirus 11	1.0x105 TCID50/mL
Bordetella bronchiseptica	1.0x106 CFU/mL
Bordetella pertussis	1.0x106 CFU/mL
Candida albicans	1.0x106 CFU/mL
Chlamydophila pneumoniae	5.0x104 TCID50/mL*
Coronavirus 229E	1.0x105 TCID50/mL
Corynebacterium diptheriae	1.0x106 CFU/mL

Coxsackievirus B5/10/2006	1.0x10⁵ TCID₅₀/mL
Echovirus 11	1.0x10⁵ TCID₅₀/mL
Echovirus 6	1.0x10⁵ TCID₅₀/mL
Enterovirus 70	1.0x10⁵ TCID₅₀/mL
Enterovirus 71	2.0x10⁴ TCID₅₀/mL*
Epstein Barr virus	1.0x10⁵ TCID₅₀/mL
Escherichia coli	1.0x10⁶ CFU/mL
Haemophilus influenzae	1.0x10⁶ CFU/mL
HSV 1 MacIntyre Strain	1.0x10⁵ TCID₅₀/mL
HSV 2 G strain	1.0x10⁵ TCID₅₀/mL
Human Rhinovirus	1.0x10⁵ TCID₅₀/mL
Klebsiella pneumoniae	1.0x10⁶ CFU/mL
Lactobacillus plantarum	1.0x10⁶ CFU/mL
Legionella pneumophila	1.0x10⁶ CFU/mL
Measles	1.0x10⁵ TCID₅₀/mL
Metapneumovirus A1	1.0x10⁵ TCID₅₀/mL
Moraxella catarrhalis	1.0x10⁶ CFU/mL
Mumps	1.0x10⁵ TCID₅₀/mL
Mycobacterium avium	1.0x10⁶ CFU/mL
Mycobacterium tuberculosis	1.0x10⁶ CFU/mL
Mycoplasma pneumoniae	1.0x10⁶ CFU/mL
Neisseria gonorrhoeae	1.0x10⁶ CFU/mL
Neisseria meningitides	1.0x10⁶ CFU/mL
Parainfluenza Type 1	1.0x10⁵ TCID₅₀/mL
Parainfluenza Type 2	1.0x10⁵ TCID₅₀/mL
Parainfluenza Type 3	1.0x10⁵ TCID₅₀/mL
Proteus mirabilis	1.0x10⁶ CFU/mL
Proteus vulgaris	1.0x10⁶ CFU/mL
Pseudomonas aeruginosa	1.0x10⁶ CFU/mL
Respiratory syncytial virus	1.0x10⁵ TCID₅₀/mL
Staphylococcus aureus	1.0x10⁶ CFU/mL
Staphylococcus epidermidis	1.0x10⁶ CFU/mL
Streptococcus mutans	1.0x10⁶ CFU/mL
Streptococcus pneumoniae	1.0x10⁵ CFU/mL*
Streptococcus pyogenes	1.0x10⁶ CFU/mL
Streptococcus salivarius	1.0x10⁶ CFU/mL

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Due to low concentration of the stock organism, the concentration tested was below the target. The actual concentration tested is listed in the table.

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No cross-reactivity was observed with the forty-four (44) microorganisms tested with the Solana® Influenza A+B Assay.

Interference:

The performance of Solana® Influenza A+B Assay was evaluated with potentially interfering substances that may be present in nasopharyngeal specimens. The potentially interfering substances were evaluated with influenza A (A/Mexico/ 4108/2009) and influenza B (Influenza B/Brisbane/60/08) at concentrations of 2x LOD. There was no evidence of interference caused by the substances tested at the concentrations shown below.

Table 8. Interfering Substances
Substances	Active Ingredient	Concentration Tested
Purified mucin protein	Mucin protein	2.5 mg/mL
Blood (human)	Blood	5.0%
Afrin — nasal spray	Oxymetazoline	5.0%
Saline nasal spray	Saline	15.0%
Phenylephrine
hydrochloride	Phenylephrine hydrochloride	15.0%
Flonase	Fluticasone	5.0%
Zicam Gentle Allergy Relief
NasalGel	Galphimia glauca, Histaminum
hydrochloricum, Luffa operculata,
Sulfur	5.0%
Mupirocin	Mupirocin	12.0 mg/mL
Oseltamivir	Oseltamivir	2.2 µg/mL
Zanamivir	Zanamivir	282.0 ng/mL
Tobramycin	Tobramycin	2.5 mg/mL
Chloraseptic	Benzocaine, Menthol	0.68 g/mL
Amantadine hydrochloride	Amantadine hydrochloride	282.0 ng/mL
Nasocort Allergy 24 hour	Triamcinolone	5.0%
Sinus Buster Nasal Spray	Capsicum annuum (Capsaicin)	5.0%
NasalCrom Nasal Allergy	Cromolyn Sodium	5.0%

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Table 8.Interfering Substances
Substances	Active Ingredient	Concentration Tested
Spray
Rhinocort	Budesonide (Glucocorticoid)	5.0%
Air-Vita Allergy Multi-
Symptom Relief	Allium cepa, Ambrosia
artemisiaefolia, Apis mellifica,
Chamomilla, Eucalyptol, Eucalyptus
globulus, Euphrasia officinalis,
Galphimia glauca, Histaminum
hydrochloricum, Natrum
muriaticum, Nux vomica, Quercus
robur, Silicea, Wyethia helenioides	5.0%
Ipratropium bromide	Ipratropium bromide	10.0 mg/mL
Olopatadine hydrochloride	Olopatadine hydrochloride	10.0 mg/mL
Amantadine hydrochloride	Amantadine hydrochloride	282.0 ng/mL

Analytical Reactivity (Inclusivity):

The reactivity of the Solana® Influenza A+B Assay was evaluated against multiple strains of influenza A and influenza B viruses. The clinical influenza panel consisted of fourteen (14) influenza A strains, and eight (8) Influenza B strains at concentrations near the level of detection (LOD) of the assay.

Table 9. Inclusivity Strains
Strain	Subtype/Lineage	TCID50/mL	Inclusive (Yes or
No)
	Influenza A
A/Mexico/4108/2009	H1N1p	2.3x103	Yes
A/Denver/1/57	H1N1	2.3x103	Yes
A/New Jersey/8/76	H1N1	2.3x103	Yes
A/PR/8/34	H1N1	2.3x103	Yes
A/FM/1/47	H1N1	2.3x103	Yes
A/Solomon Islands/3/06	H1N1	2.3x103	Yes
A/New Caledonia/20/1999	H1N1	2.3x103	Yes
A/Victoria/361/11	H3N2	2.3x103	Yes
A/Port Chalmers/1/73	H3N2	1.4x104	Yes
A/Aichi/2/68	H3N2	2.3x103	Yes
A/Victoria/3/75	H3N2	2.3x103	Yes

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Table 9.Inclusivity Strains
Strain	Subtype/Lineage	TCID50/mL	Inclusive (Yes or
No)
A/Hong Kong/8/68	H3N2	2.3x103	Yes
A/Wisconsin/67/2005	H3N2	2.3x103	Yes
A/WS/33	H1N1	2.3x103	Yes
	Influenza B
B/Malaysia/2506/04	Victoria	2.6x102	Yes
B/Florida/07/2004	Victoria	7.7x102	Yes
B/Maryland/1/59	Yamagata	2.6x102	Yes
B/Allen/45	Yamagata	2.6x102	Yes
B/Lee/40	Yamagata	2.6x102	Yes
B/Florida/04/2006	Yamagata	7.7x102	Yes
B/Panama/45/90	Yamagata	2.6x102	Yes
B/Hong Kong/5/72	Victoria	2.6x102	Yes
B/Malaysia/25/06/04	Victoria	2.6x102	Yes

Due to restrictions and availability of a number of influenza A strains, in silico analysis was performed for three additional strain designations:

A total of four (4) H3N2v (1 human strain and 3 swine) sequences were analyzed in silico. All four sequences demonstrated 100% homology.
. A total of three hundred forty (340) H5N1 strains were analyzed in silico. Three hundred thirty-nine (339) strains in the database demonstrated ≥95% overall homology and ≥88% homology to any individual primer or probe sequence. One H5N1 strain demonstrated an overall homology of 88% and ≥82% homology to any individual primer or probe sequence.
A total of one hundred sixty-four (164) H7N9 sequences were analyzed in silico. All 164 sequences demonstrated 100% homology.
Fourteen (14) non-clinical avian restricted influenza A viruses (Table 10) were analyzed in silico.

Table 10.	Non-clinical Avian Restricted Influenza A Viruses
Subtype	Strain
H2N2	A/Mallard/NY/6750/78 (H2N2)
H7N3	A/Chicken/NJ/15086-3/94 (H7N3)
H9N2	A/Chicken/NJ/12220/97 (H9N2)
H4N8	A/Mallard/OH/338/86 (H4N8)
H6N2	A/Chicken/CA/431/00 (H6N2)

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Table 10.	Non-clinical Avian Restricted Influenza A Viruses
Subtype	Strain
H8N4	A/Blue Winged Teal/LA/B174/86 (H8N4)
H5N1	A/Anhui/01/2005(H5N1)-PR8-IBCDC-RG5
H10N7	A/GWT/LA/169GW/88 (H10N7)
H11N9	A/Chicken/NJ/15906-9/96 (H11N9)
H12N5	A/Duck/LA/188D/87 (H12N5)
H13N6	A/Gull/MD/704/77 (H13N6)
H14N5	A/Mallard/GurjevRussia/262/82 (H14N5)
H15N9	A/Shearwater/Australia/2576/79 (H15N9)
H16N3	A/Shorebird/DE/172/2006(H16N3)

A total of twenty-seven (27) sequences were available for analysis. The Solana™ FluA primers and probe are 90-100% conserved to the specified avian strains and to representative avian strains.

f. Assay cut-off:
Not applicable.

2. Comparison studies:

a. Method comparison with predicate device:
Not applicable
b. Matrix comparison:
Not applicable
1. Clinical studies:
- a. Clinical Sensitivity:

22

Performance characteristics of the Solana Influenza A+B Assay were established during a prospective study with specimens collected between February and April 2016. One thousand four hundred seventy-three (1473) prospectively collected specimens have been included in this study at five (5) sites across the United States. A single nasal or nasopharyngeal swab specimen (302 and 1171, respectively) was collected per patient in viral transport media (BD/Copan UTM, Remel M5, Remel M6). All specimens were transported to a central location at 2℃ to 8℃ for testing by the comparator methods (culture for influenza A and B using the R-Mix Too mixed cells and direct specimen DFA (DSFA), and extraction with the NucliSENS® easyMAG® and testing with a FDA-cleared Influenza A+B molecular assay). The specimens were processed and tested with Solana Influenza A+B Assay (frozen (731) and fresh (742)) on the Solana instrument at the sites.

COMPARISON VERSUS CULTURE WITH DFA AND DSFA

One thousand four hundred seventy-three (1473) fresh specimens were included in this study. Each specimen was cultured for influenza A and B using the R-Mix Too mixed cells and stained with an FDA-cleared device and processed for direct specimen DFA (DSFA). All comparator testing was performed on fresh specimens within 72-hours of their collection. A specimen was recorded as positive for influenza A or B if either comparator test was positive.

Seven hundred and forty-two (742) of these specimens were tested fresh using the Solana Influenza A+B Assay for the presence influenza A or B. Seven hundred and thirty-one (731) specimens were frozen and stored at -70°C prior to testing with the Solana Influenza A+B Assay. Fifteen (15) specimens were contaminated or toxic in the cell culture (1.0%). Fifty (50) specimens were invalid in the Solana Assay (3.4%). These sixty-five (65) specimens have been excluded from further analysis. Tables 11 and 12 detail the performance of the Solana Assay for influenza A and influenza B respectively for the remaining one thousand four hundred eight (1408) specimens, across all testing sites combined, as compared to viral culture with DSFA results.

| Table 11. | | Performance Characteristics of the Solana Influenza A+B Assay for Influenza A
Compared to Culture and DSFA (Across all Sites Combined) | | | | | |
|--------------------|------|-------------------------------------------------------------------------------------------------------------------------------------------|-----|-----|-----|--------------------------|--------------------------|
| Source
Category | N | TP | FP | TN | FN | Sensitivity%
(95% CI) | Specificity%
(95% CI) |
| Fresh | 709 | 180 | 24 | 503 | 2 | 98.9
(96.1 to 99.7) | 95.4
(93.3 to 96.9) |
| Frozen | 699 | 176 | 27 | 493 | 3 | 98.3
(95.2 to 99.4) | 94.8
(92.6 to 96.4) |
| All | 1408 | 356 | 51* | 996 | 5** | 98.6
(96.8 to 99.4) | 95.1
(93.7 to 96.3) |

*Of the fifty-one (51) discordant specimens (Solana Positive/Culture and DSFA Negative) reported in Table 11, twenty-eight (28) of these specimens were positive by an alternate molecular assay.

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| Table 12. | | Performance Characteristics of the Solana Influenza A+B Assay for Influenza B Compared to
Culture and DSFA (Across all Sites Combined) | | | | | |
|--------------------|------|-------------------------------------------------------------------------------------------------------------------------------------------|----|------|----|--------------------------|--------------------------|
| Source
Category | N | TP | FP | TN | FN | Sensitivity%
(95% CI) | Specificity%
(95% CI) |
| Fresh | 709 | 62 | 1 | 646 | 0 | 100
(94.2 to 100) | 99.8
(99.1 to 100) |
| Frozen | 699 | 23 | 8 | 668 | 0 | 100
(85.7 to 100) | 98.8
(97.7 to 99.4) |
| All | 1408 | 85 | 9* | 1314 | 0 | 100
(95.7 to 100) | 99.3
(98.7 to 99.6) |

**Of the five (5) discordant specimens (Solana Negative/Culture and DSFA Positive) reported in Table 11, two (2) of these specimens were positive by an alternate FDA cleared molecular assay.

*Of the nine (9) discordant specimens (Solana Positive/Culture and DSFA Negative) reported in Table 12, two (2) of these specimens were positive by an alternate FDA cleared molecular assay.

COMPARISON WITH A FDA-CLEARED INFLUENZA A+B MOLECULAR ASSAY

One thousand four hundred seventy-three (1473) specimens were processed using the NucliSENS® easyMAG® and tested with a FDA-cleared Influenza A+B molecular assay according to the assay's package insert. The comparator testing was performed on fresh specimens within 72-hours of their collection.

Seven hundred and thirty-one (731) of the original specimens were frozen and stored at -70°C prior to testing with the Solana Influenza A+B Assay. Seven hundred and forty-two (742) of the original specimens were tested fresh using the Solana Influenza A+B Assay for the presence of influenza A or B. Thirty one (31) specimens were invalid in the comparator assay (2.1%). Fifty (50) specimens were invalid in the Solana Assay (3.4%)(one specimen was invalid in both assays). These eighty (80) specimens have been excluded from further analysis.

Tables 13 and 14 detail the positive percent agreement (PPA) and the negative percent agreement (NPA) of the Solana Influenza A+B Assay results for influenza A and influenza B respectively, as compared with an FDA cleared molecular comparator, for the remaining one thousand three hundred ninety-three (1393) specimens.

| Table 13.
Percent Agreement of the Solana® Influenza A+B Assay for Influenza A

Compared to an FDA cleared Influenza A+B Molecular Assay (Across all Sites Combined)
Source
Category	N	TP	FP	TN	FN	PPA
(95% CI)	NPA
(95% CI)
Fresh	710	195	9	499	7	96.5
(93.0 to 98.3)	98.2
(98.7 to 99.1)
Frozen	683	180	24	475	4	97.8
(94.5 to 99.2)	95.2
(92.9 to 96.7)

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Table 13. Percent Agreement of the Solana® Influenza A+B Assay for Influenza A Compared to an FDA cleared Influenza A+B Molecular Assay (Across all Sites Combined)
Source Category	N	TP	FP	TN	FN	PPA (95% CI)	NPA (95% CI)
All	1393	375	33	974	11	97.2 (95.0 to 98.4)	96.7 (95.4 to 97.7)

There were a total of forty-four (44) discordant specimens among the hundred ninety-three (1393) specimens evaluated. Of the thirty-three (33) discordant specimens (Solana Positive) Comparator Negative) reported in Table 13, nine (9) of these specimens were positive by culture/DSFA. Of the eleven (11) discordant specimens (Solana Negative/Comparator Positive) reported in Table 13, two (2) of these specimens were positive by culture/DSFA.

| Percent Agreement of the Solana® Influenza A+B Assay for Influenza B

Compared to an FDA-cleared Influenza A+B Molecular Assay (Across all Sites Combined)
Table 14.	Source
Category	N	TP	FP	TN	FN	PPA
(95% CI)	NPA
(95% CI)
	Fresh	710	57	6	647	0	100
(93.7 to 100)	99.1
(98.0 to 99.6)
	Frozen	683	23	8	652	0	100
(85.7 to 100)	98.8
(97.6 to 99.4)
	All	1393	80	14	1299	0	100
(95.4 to 100)	98.9
(98.2 to 99.4)

There were a total of fourteen (14) discordant specimens among the one thousand three hundred ninety-three (1393) specimens evaluated. Of the fourteen (14) discordant specimens (Solana Positive) Comparator Negative) reported in Table 14, seven (7) of these specimens were positive by culture/DSFA.

b. Clinical specificity:

See Section 3a.

Other clinical supportive data (when a. and b. are not applicable): C.

Not applicable

1. Clinical cut-off:
  Not applicable
1. Expected values:
  The expected values of the Solana Influenza A+B Assay were established during a prospective study conducted between February and April 2016. One thousand four hundred seventy-three (1473) specimens (fresh (742) and frozen (731)) have been included in this study at five (5) sites across the United States. A single specimen was

25

collected per patient. The specimens were processed and tested with Solana® Influenza A+B Assay on the Solana® instrument at the sites.

The expected value of influenza A and influenza B with the Solana Influenza A+B Assay has been calculated for the combined sites based on the age of the patient.

Fifty-three (53) of the one thousand four hundred seventy-three (1473) specimens were removed from analysis: (three (3) specimens did not have the age provided; fifty (50) specimens were invalid). Table 15 provides the percentage of influenza A and influenza B positive cases per specified age group, as determined by the Solana Influenza A+B Assay, for the remaining one thousand four hundred twenty (1420) specimens.

Table 15. Expected Values (N=1420)
Age Group	Influenza A			Influenza B
	Number of Patients	Number of Positives	Prevalence	Number of Patients	Number of Positives	Prevalence
≤ 5 years	377	91	24.1%	377	26	6.9%
6 to 21 years	297	89	30.0%	297	48	16.2%
22 to 59 years	504	191	37.9%	504	17	3.4%
≥ 60 years	242	37	15.3%	242	3	1.2%

The prospective clinical study had a dual infection rate for Influenza A and Influenza B of 0.2% (3/1420) using the Solana Influenza A+B Assay. All three (3) of these dual detections were only positive for influenza A by culture and DSFA and also by an alternate molecular comparator.

N. Other Supportive Instrument Performance Characteristics Data Not Covered In The "Performance Characteristics" Section above:

Instrument: Solana Instrument

O. System Descriptions:

1. Modes of Operation:

The Solana instrument heats each reaction tube to 58°C. If present, the target RNA sequence is reverse transcribed into cDNA by the Reverse Transcriptase and influenza A or B specific primers that are present in the reaction mix. After the completion of the Reverse Transcriptase step, the Solana instrument heats each reaction tube to 65°C where the isothermal DNA polymerase amplifies the cDNA strands using the influenza A or B specific primers. Influenza A or B specific fluorescence probes are also included in the Reaction Tube. The target probes are labeled with a quencher on one end and a fluorophore specific for the recognition sequence on the other end. In addition, the

26

target probes carry a ribonucleic acid. Upon annealing to amplicons, the fluorescence probes are cleaved by RNaseH2 and the fluorescence signal increases due to physical separation of fluorophore from quencher. The Solana instrument measures and interprets the fluorescent signal, using on-board method-specific algorithms. Solana instrument will then report the test results to the user on its display screen, and it can print out the results via a printer.

2. Software:

FDA has reviewed applicant's Hazard Analysis and software development processes for this line of product types:

Yes No

P. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10, 21 CFR 801.109, and the special controls.