JBAIDS Influenza A & B Detection Kit

K152523 · U.S Army Medical Materiel Development Activity · OCC · Oct 1, 2015 · Microbiology

Device Facts

• Record ID: K152523
• Device Name: JBAIDS Influenza A & B Detection Kit
• Applicant: U.S Army Medical Materiel Development Activity
• Product Code: OCC · Microbiology
• Decision Date: Oct 1, 2015
• Decision: SESE
• Submission Type: Special
• Regulation: 21 CFR 866.3980
• Device Class: Class 2

Intended Use

The Joint Biological Agent Identification and Diagnostic System (JBAIDS) Influenza A & B Detection Kit is intended for use on the JBAIDS instruments, for the in vitro qualitative detection of influenza A and influenza B viral nucleic acids isolated and purified nasopharyngeal swab (NPS) and nasopharyngeal wash (NPW) specimens from human patients with signs and symptoms of respiratory infection. The JBAIDS Influenza A & B Detection Kit contains reverse transcriptase real-time polymerase chain reaction (rRT-PCR) assays that target the Matrix protein gene of influenza A viruses, and the Non-structural protein gene of influenza B viruses. This kit is not intended to detect influenza C viruses. Test results are to be used in conjunction with other clinical and epidemiological information. Negative results do not preclude influenza virus infection and should not be used as the sole basis for treatment or other patient management decisions. Performance characteristics for detection of influenza A were established when 2009 H1N1 Influenza, Influenza A H1N1, and Influenza A H3N2 were the predominant influenza A viruses in circulation. Due to low seasonal prevalence, performance characteristics for detection of seasonal Influenza A/H1 were established primarily with retrospective and surrogate clinical specimens. When other influenza A viruses are present, performance characteristics may vary. All users, analysts, and any person reporting diagnostic results from use of this device should be trained to perform and interpret the results from this procedure by JBAIDS instructors or designees prior to use. Use of this device is limited to designated Department of Defense (DoD) laboratories equipped with the JBAIDS instruments. 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

Device Story

JBAIDS Influenza A & B Detection Kit uses rRT-PCR to detect viral nucleic acids; targets Matrix protein gene (Influenza A) and Non-structural protein gene (Influenza B). Input: purified nucleic acids from nasopharyngeal swabs/washes. Operation: rRT-PCR performed on JBAIDS instruments in designated DoD laboratories by trained personnel. Output: qualitative detection of Influenza A and B. Modification: updated analytical inclusivity to include 5 specific influenza A strains (H7N9, H3N2, H3N2v). Clinical utility: results used alongside clinical/epidemiological data to support patient management; negative results do not rule out infection. Benefit: provides rapid identification of influenza strains in military/DoD settings.

Clinical Evidence

Bench testing only. Analytical sensitivity (Limit of Detection) established for H7N9 and H3N2v strains using simulated nasopharyngeal swab samples. LoD for H7N9 determined to be 4 x 10^4 EID50/mL; LoD for H3N2v strains ranged from 2.0 to 31.6 TCID50/mL. No clinical prospective or retrospective patient data provided in this submission.

Technological Characteristics

rRT-PCR assay; hydrolysis probe chemistry (6-FAM/TAMRA). Targets: Influenza A matrix gene, Influenza B non-structural gene, RNAse P (control). Form factor: freeze-dried reagent vials. Instrumentation: JBAIDS instrument. Extraction: IT 1-2-3 Platinum Path or Roche MagNA Pure Compact. Connectivity: JBAIDS instrument-integrated software. Storage: room temperature.

Indications for Use

Indicated for in vitro qualitative detection of influenza A and B viral nucleic acids in nasopharyngeal swab and wash specimens from human patients with respiratory infection symptoms.

Regulatory Classification

Identification

A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B; (2) Influenza A subtype H1 and Influenza A subtype H3; (3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B; (4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus; (5) Human Metapneumovirus; (6) Rhinovirus; and (7) Adenovirus.

Special Controls

*Classification.* Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;” (2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and (3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.

Predicate Devices

• JBAIDS Influenza A & B Detection Kit (K111775)

Submission Summary (Full Text)

{0}------------------------------------------------

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized depiction of an eagle or bird-like figure with three overlapping lines forming its body and wings. The logo is encircled by the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" in a circular arrangement.

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

October 1, 2015

U.S. Army Medical Materiel Development Activity Robert Miller, PhD, RAC Director, Division of Regulated Activities and Compliance 1430 Veterans Drive Fort Detrick, MD 21702-9232

Re: K152523

Trade/Device Name: JBAIDS Influenza A & B Detection Kit Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: II Product Code: OCC Dated: September 2, 2015 Received: September 3, 2015

Dear Dr. Miller:

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. 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 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.

{1}------------------------------------------------

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. 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 Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

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

Sincerely yours,

# Tamara V. Feldblyum -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) K152523

Device Name JBAIDS Influenza A & B Detection Kit

## Indications for Use (Describe)

The Joint Biological Agent Identification and Diagnostic System (JBAIDS) Influenza A & B Detection Kit is intended for use on the JBAIDS instruments, for the in vitro qualitative detection of influenza B viral nucleic acids isolated and purified NPS and NPW specimens from human patients with signs and symptoms of respiratory infection.

The JBAIDS Influenza A & B Detection Kit contains reverse transcriptase real-time polymerase chain reaction (rRT-PCR) assays that target the Matrix protein gene of influenza A viruses, and the Non-structural protein gene of influenza B viruses. This kit is not intended to detect influenza C viruses.

Test results are to be used in conjunction with other clinical and epidemiological information. Negative results do not preclude influenza virus infection and should not be used as the sole basis for treatment or other patient management decisions.

Performance characteristics for detection of influenza A were established when 2009 H1N1 Influenza, Influenza A H1N1, and Influenza A H3N2 were the predominant influenza A viruses in circulation. Due to low seasonal prevalence, performance characteristics for detection of seasonal Influenza A/H1 were established primarily with retrospective and surrogate clinical specimens. When other influenza A viruses are present, performance characteristics may vary. All users, and any person reporting diagnostic results from use of this device should be trained to perform and interpret the results from this procedure by JBAIDS instructors or designees prior to use. Use of this device is limited to designated Department of Defense (DoD) laboratories equipped with the JBAIDS instruments.

If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a biosatory (BSL) 3+ facility is available to receive and culture specimens.

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

|X | Prescription Use (Part 21 CFR 801 Subpart D)

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

CONTINUE ON A SEPARATE PAGE IF NEEDED.
This section applies only to requirements of the Paperwork Reduction Act of 1995.

| *DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.* | |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--|
| The burden time for this collection of information is estimated to average 79 hours per response, including the<br>time to review instructions, search existing data sources, gather and maintain the data needed and complete<br>and review the collection of information. Send comments regarding this burden estimate or any other aspect<br>of this information collection, including suggestions for reducing this burden, to: | |
| Department of Health and Human Services<br>Food and Drug Administration<br>Office of Chief Information Officer<br>Paperwork Reduction Act (PRA) Staff<br>[e-mail] | |
| "An agency may not conduct or sponsor, and a person is not required to respond to, a collection of | |

information unless it displays a currently valid OMB number."

{3}------------------------------------------------

Image /page/3/Picture/0 description: The image is a black and white seal for the Department of Defense. The seal is circular and contains an eagle in the center with its wings spread. The eagle is holding arrows and an olive branch. Above the eagle are 13 stars arranged in a semi-circle. The words "DEPARTMENT OF DEFENSE" are written around the top of the circle, and the words "UNITED STATES OF AMERICA" are written around the bottom of the circle.

DEPARTMENT OF THE A U.S. ARMY MEDICAL MATERIEL DEVELOPMENT ACTIVITY 1430 VETERANS DRIVE FORT DETRICK, MARYLAND 21702-5009

# 510(k) Summary

#### 1. Introduction

According to the requirements of 21 CFR 807.92, the following information provides sufficient detail to understand the basis for a determination of substantial equivalence.

| 2.<br>Administrative Information | |
|----------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Submitted by: | The Surgeon General, Department of the Army |
| Primary Contact: | Robert Miller, PhD, RAC<br>Director, Division of Regulated Activities and Compliance<br>Telephone: [phone]<br>Fax: [phone]<br>Email: usarmy.detrick.medcom-usammda.mbx.usamrmc-<br>[e-mail] |
| Secondary Contact: | Chandar Thakur, PhD, RAC<br>Regulatory Affairs Scientist, Division of Regulated Activities and<br>Compliance<br>Telephone: [phone]<br>Fax: [phone]<br>Email: [e-mail] or usarmy.detrick.medcom-<br>[e-mail] |
| Technical Contact: | Cynthia Phillips, PhD<br>Director, Regulated Products, BioFire Defense, LLC<br>Telephone:[phone], ext 1370<br>Fax: [phone]<br>Email: [e-mail] |
| 3.<br>Device | |
| Device Name/ Trade Name: | JBAIDS Influenza A & B Detection Kit |
| Common Name: | Real-time PCR assay for detection of Influenza A and Influenza B |
| Classification Name: | Respiratory viral panel multiplex nucleic acid assay (21 CFR<br>866.3980) |

{4}------------------------------------------------

Image /page/4/Picture/0 description: The image is the seal of the Department of Defense. The seal is circular and contains an eagle with its wings spread, holding arrows and an olive branch. Above the eagle are 13 stars arranged in a semi-circle. The words "DEPARTMENT OF DEFENSE" are written around the top of the seal, and "UNITED STATES OF AMERICA" are written around the bottom.

DEPARTMENT OF THE AR U.S. ARMY MEDICAL MATERIEL DEVELOPMENT 1430 VETERANS DRIVE FORT DETRICK, MARYLAND 21702-500

#### 4. Intended Use

The Joint Biological Agent Identification and Diagnostic System (JBAIDS) Influenza A & B Detection Kit is intended for use on the JBAIDS instruments, for the in vitro qualitative detection of influenza A and influenza B viral nucleic acids isolated and purified nasopharygeal swab (NPS) and nasopharygeal wash (NPW) specimens from human patients with signs and symptoms of respiratory infection.

The JBAIDS Influenza A & B Detection Kit contains reverse transcriptase real-time polymerase chain reaction (rRT-PCR) assays that target the Matrix protein gene of influenza A viruses, and the Non-structural protein gene of influenza B viruses. This kit is not intended to detect influenza C viruses.

Test results are to be used in conjunction with other clinical and epidemiological information. Negative results do not preclude influenza virus infection and should not be used as the sole basis for treatment or other patient management decisions.

Performance characteristics for detection of influenza A were established when 2009 H1N1 Influenza, Influenza A H1N1, and Influenza A H3N2 were the predominant influenza A viruses in circulation. Due to low seasonal prevalence, performance characteristics for detection of seasonal Influenza A/H1 were established primarily with retrospective and surrogate clinical specimens. When other influenza A viruses are present, performance characteristics may vary.

All users, analysts, and any person reporting diagnostic results from use of this device should be trained to perform and interpret the results from this procedure by JBAIDS instructors or designees prior to use. Use of this device is limited to designated Department of Defense (DoD) laboratories equipped with the JBAIDS instruments.

If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a biosafety (BSL) 3+ facility is available to receive and culture specimens.

#### 5. Device Description

The JBAIDS Influenza A & B Detection Kit is a rRT-PCR test kit, which, when used with the JBAIDS instrument and software, allows the qualitative in vitro detection of influenza A and B viral RNA. These two assays have been optimized as freeze-dried assays with primer and fluorescent-probe sets for the detection of influenza A and B viral RNA. In particular, the influenza A assay targets a region of the matrix gene specific to the influenza A virus genera, and the influenza B assay targets a region of the nonstructural gene specific to the influenza B virus genera. The tests are performed using the previously FDA-cleared JBAIDS instrument and software. A human gene target assay (RNAse P target) will be used as an inhibition and extraction control.

{5}------------------------------------------------

Image /page/5/Picture/0 description: The image is a black and white seal for the Department of Defense. The seal is circular and contains an eagle with its wings spread, clutching arrows and an olive branch. Above the eagle are 13 stars arranged in an arc. The words "DEPARTMENT OF DEFENSE" are written around the top of the circle, and "UNITED STATES OF AMERICA" is written around the bottom.

DEPARTMENT OF THE AR U.S. ARMY MEDICAL MATERIEL DEVELOPMENT 1430 VETERANS DRIVE FORT DETRICK, MARYLAND 21702-5009

#### 6. Assay Principle

Before testing, NPS or NPW specimens are purified using Idaho Technology's 1-2-3TM Platinum Path Sample Purification Kit or the Roche MagNA Pure Compact Nucleic Acid Isolation Kit I. The resulting purified sample is added to unknown reagent vials and a sample control reagent vial, along with reconstitution buffer. When viral RNA is present, a fragment of influenza A or B viral RNA is transcribed and amplified. The amplicon is detected by fluorescence using a specific hydrolysis probe. Each probe is labeled on one end with a fluorescent reporter moiety (6-carboxyfluorescein (6-FAM)) and elsewhere with a quencher moiety (carboxy tetramethylrhodamine (TAMRA)). When the probe is intact, the quencher absorbs the light emitted by the reporter moiety. During PCR, the probe hybridizes to the target sequence before the exonuclease activity of Taq polymerase hydrolyzes the probe, separating the fluorophore from the quencher and permitting detection of the fluorescent signal generated by the reporter. The fluorescent signal increases as additional templates are amplified and more probes are hydrolyzed. JBAIDS Software analyzes the fluorescence amplification curves and reports results as positive, negative, or uncertain. A failure of the positive control will result in the entire run being called invalid. Failure of the sample control vields a result of "sample control failure" when the associated sample has a negative result for the target assay. Retesting is required to resolve uncertain, invalid, or sample control failure results.

#### 7. Substantial Equivalence

The modified JBAIDS Influenza A & B Detection System is substantially equivalent to the current legally marketed device. JBAIDS Influenza A & B Detection Kit. Additions made to the labeling to add additional strain testing did not change the intended use of the device or the fundamental scientific technology.

The JBAIDS Influenza A & B Detection Kit with additional labeling for detection of Influenza A H3N2v and H7N9 strains described is substantially equivalent to the JBAIDS Influenza A & B Detection Kit, which was cleared on September 13, 2011 under 510(k) K111775.

{6}------------------------------------------------

Image /page/6/Picture/0 description: The image is a black and white seal for the Department of Defense. The seal is circular and contains an eagle with its wings spread, holding arrows and an olive branch. Above the eagle are 13 stars arranged in a semi-circle. The words "DEPARTMENT OF DEFENSE" are written around the top of the circle, and "UNITED STATES OF AMERICA" are written around the bottom.

### DEPARTMENT OF THE ARN U.S. ARMY MEDICAL MATERIEL DEVELOPMENT ACTIVITY 1430 VETERANS DRIVE FORT DETRICK, MARYLAND 21702-5009

### Table 1: Similarities Between the JBAIDS Influenza A & B Detection Kit (this submission) and the JBAIDS Influenza A & B Detection Kit (K111775)

| Element | JBAIDS Influenza A and B Detection kit<br>(this submission) | JBAIDS Influenza A and B<br>Detection kit (K111775) |
|-----------------------------------|---------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------|
| Technology | Real time PCR using hydrolysis probes | Same |
| Viruses Detected | Qualitative <i>in vitro</i> detection of Influenza<br>A and Influenza B virus nucleic acids | Same (see Table 2) |
| Specimen Types | Nasopharyngeal swabs and<br>Nasopharyngeal washes | Same (see Table 2) |
| Extraction Methods | IT 1-2-3TM Platinum Path Sample<br>Purification Kit and Roche MagNA Pure<br>Compact Nucleic Acid Isolation<br>Kit I | Same (see Table 2) |
| Required Instrumentation | JBAIDS Instrument | Same |
| Interpretation of Test<br>Results | Automated analysis of test results and<br>controls | Same |
| Reagent Storage | Reagents are stored at room temperature | Same |

#### Table 2: Differences Between the JBAIDS Influenza A & B Detection Kit (this submission) and the JBAIDS Influenza A & B Detection Kit (K111775)

| Element | JBAIDS Influenza A and B Detection kit<br>(this submission) | JBAIDS Influenza A and B<br>Detection kit (K111775) |
|--------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------|
| Organisms Detected | Demonstrated inclusive detection of<br>Influenza A H3N2v and A H7N9 as<br>positive for Influenza A | Not labeled for detection of<br>Influenza A H3N2v and H7N9. |
| Specimen Types | Demonstrated detection of A H3N2v and A<br>H7N9 from simulated nasopharyngeal<br>swabs. No testing of these strains has been<br>performed on nasopharyngeal washes. | Nasopharyngeal swabs and<br>washes |
| Extraction methods | Demonstrated detection of Influenza A<br>H3N2v and A H7N9 from simulated<br>nasopharyngeal swab samples extracted<br>using the IT 1-2-3TM Platinum Path Sample<br>Purification Kit. No testing of these strains<br>was performed on samples extracted using<br>the Roche MagNA Pure Compact Nucleic<br>Acid Isolation Kit. | IT 1-2-3TM Platinum Path Sample<br>Purification Kit and Roche MagNA<br>Pure Compact Nucleic Acid<br>Isolation Kit |

#### 8. Selected Analytical Studies

The estimated analytical sensitivity or Limit of Detection (LoD) was determined for four strains of A H3N2 (two seasonal A H3N2 strains, and two swine variant, or A H3N2v, strains) and one strain of H7N9 using the Flu A assay. Simulated nasopharyngeal swab samples were extracted using the IT 1-2-3™ Platinum Path Sample Purification Kit prior to testing with the JBAIDS

{7}------------------------------------------------

Image /page/7/Picture/0 description: The image is a black and white seal for the Department of Defense. The seal is circular and contains an eagle with outstretched wings in the center. Above the eagle are 13 stars arranged in a semi-circle. The words "DEPARTMENT OF DEFENSE" are written around the top of the circle, and "UNITED STATES OF AMERICA" are written around the bottom.

DEPARTMENT OF THE A U.S. ARMY MEDICAL MATERIEL DEVELOPMENT ACTIVITY 1430 VETERANS DRIVE FORT DETRICK, MARYLAND 21702-5009

Influenza A & B Detection kit. Serial dilutions were run in triplicate, with all three replicates required to read positive in order for that dilution to be called positive. Results are shown in Table 3 and Table 4. This data was added to the analytical inclusivity section of the package insert.

Estimation studies suggested that the LOD for H7N9 was 4 x 104 EID50 /mL. Upon refinement and running further replicates, the confirmed LOD for H7N9 was determined to be 4 x 10 EIDs0 /mL. LOD estimation studies suggested that the LOD for H3N2v was between 2 and 32 TCID-0 /mL.

| Strain | Concentration | No. Positive<br>Replicates |
|----------------|--------------------|----------------------------|
| A/Anhui/1/2013 | 8.0 x 105 EID50/ml | 3/3 |
| A/Anhui/1/2013 | 4.0 x 105 EID50/ml | 3/3 |
| A/Anhui/1/2013 | 8.0 x 104 EID50/ml | 3/3 |
| A/Anhui/1/2013 | 4.0 x 104 EID50/ml | 3/3 |
| A/Anhui/1/2013 | 8.0 x 103 EID50/ml | 2/3 |
| A/Anhui/1/2013 | 4.0 x 103 EID50/ml | 2/3 |
| A/Anhui/1/2013 | 8.0 x 102 EID50/ml | 1/3 |
| A/Anhui/1/2013 | 4.0 x 102 EID50/ml | 0/3 |
| A/Anhui/1/2013 | 8.0 x 101 EID50/ml | 2/3 |
| A/Anhui/1/2013 | 4.0 x 101 EID50/ml | 0/3 |

#### Table 3: Estimated LoD of H7N9 (A/Anhui/1/2013)

#### Estimated LoD of H3N2 and H3N2v Strains Table 4:

| Strain | Isolate | LoD (TCID50/mL) |
|---------------------------------|-------------------------|-----------------|
| Influenza A H3N2 (seasonal) | A/Victoria/361/2011 | 63.0 |
| Influenza A H3N2 (seasonal) | A/Perth/16/2009 | 6.3 |
| Influenza A swine variant H3N2v | A/West/Virginia/06/2011 | 31.6 |
| Influenza A swine variant H3N2v | A/Minnesota/11/2010 | 2.0 |

{8}------------------------------------------------

Image /page/8/Picture/0 description: The image is a black and white seal for the Department of Defense. The seal is circular and contains an eagle holding arrows and an olive branch. Above the eagle are 13 stars arranged in a semi-circle. The words "DEPARTMENT OF DEFENSE" are written along the top of the circle, and "UNITED STATES OF AMERICA" are written along the bottom.

#### 9. References

Centers for Disease Control and Prevention. Influenza A (H3N2) Variant Virus. Accessed: 26 Aug 2015: Available from: http://www.cdc.gov/flu/swineflu/h3n2v-cases.htm.

Clinical and Laboratory Standards Institute. Protocols for Determination of Limits of Detection and Limits of Quantitation. CLSI Approved Guidance EP17-A (2004).

Clinical and Laboratory Standards Institute. Evaluation of Precision Performance of Quantitative Measurements Methods; Approved Guidance-Second Edition, CLSI Approved Guidance EP5-A2 (August 2004).

United States Food and Drug Administration. In Vitro Diagnostic Devices to Detect Influenza A Viruses: Labeling and Regulatory Path, Guidance for Industry and FDA Staff (May 1, 2007).