The Joint Biological Agent Identification and Diagnostic System (JBAIDS) Influenza A Subtyping Kit is intended for the in vitro qualitative detection and differentiation of seasonal Influenza A/H1, seasonal Influenza A/H3, and 2009 H1N1Influenza viral nucleic acids isolated and purified from nasopharyngeal swab (NPS) and nasopharyngeal wash (NPW) specimens from human patients with signs and symptoms of respiratory infection, in conjunction with clinical and epidemiological risk factors. The JBAIDS Influenza A Subtyping Kit contains reverse transcriptase real-time polymerase chain reaction (rRT-PCR) assays for use on the JBAIDS instruments. The Flu A H1. Flu A H3. and Flu A H1 2009 assays of the JBAIDS Influenza A Subtyping Kit target a region of the hemagglutinin (HA) gene of the respective Influenza A virus. The Flu A Sw assay of the JBAIDS Influenza A Subtyping Kit targets a region of the nucleocapsid protein (NP) gene of the 2009 H1N1 Influenza virus, as well as some other Influenza A viruses of swine lineage. This kit is not intended to detect Influenza B or Influenza C viruses.

A negative result for all assays in the JBAIDS Influenza A Subtyping Kit is a presumptive negative result for Influenza A. These results should be confirmed using the JBAIDS Influenza A & B Detection Kit.

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.

Due to low seasonal prevalence, performance characteristics for detection of seasonal Influenza A/H1 were established primarily with retrospective and contrived clinical specimens.

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 laboratory (BSL) 3+ facility is available to receive and culture specimens.

Device Description

The JBAIDS Influenza A Subtyping Kit is a real time RT-PCR test kit, which, when used with the JBAIDS instrument and software, allows the qualitative in vitro detection and identification of influenza A subtypes H1, H3, and H1 2009 (swine lineage) viral RNA. The assays have been optimized as freeze-dried assays with primer and fluorescent-probe sets for the detection of influenza A/H1, A/H3, and A/2009 H1 viral RNA. In particular, the Flu A H1, Flu A H3, and Flu A H1 2009 assays target distinct regions of the hemagglutinin gene specific to those subtypes, and the Flu A Sw assay targets a region of the nucleocapsid protein gene as a secondary target for the influenza A/2009 H1(swine lineage) virus. The tests are performed using the JBAIDS instrument and software.

AI/ML Overview

Here's an analysis of the acceptance criteria and study details for the JBAIDS Influenza A Subtyping Kit based on the provided text:

Preamble Regarding Acceptance Criteria:
The document provided, a 510(k) Summary, details performance data but does not explicitly state pre-defined "acceptance criteria" in the format of pass/fail thresholds for the clinical study. Instead, it presents the achieved performance metrics (PPA and NPA with 95% Confidence Intervals) from the clinical and analytical studies, implicitly demonstrating that these achieved results were deemed sufficient for substantial equivalence. For the purpose of this response, I will present the reported performance as the "acceptance criteria met," recognizing that the specific numerical targets for acceptance are not explicitly listed in this summary.

1. Table of Acceptance Criteria and Reported Device Performance

As described above, explicit acceptance criteria (i.e., specific numerical targets prior to testing) were not provided in the 510(k) summary. However, the reported performance from the clinical studies served as the basis for the device's clearance. The table below summarizes the reported clinical performance.

Influenza A Strain	Sample Type	Performance Metric	Reported Performance (PPA/NPA)	95% Confidence Interval
2009 H1N1	NPW	PPA	100.0% (66/66)	94.6-100%
	NPW	NPA	99.3% (414/417)	97.9-99.9%
	NPS	PPA	100.0% (34/34)	89.7-100%
	NPS	NPA	99.6% (277/278)	98.0-100%
Seasonal H3	NPW	PPA	100.0% (33/33)	89.4-100%
	NPW	NPA	100.0% (450/450)	99.2-100%
	NPS	PPA	100.0% (26/26)	86.8-100%
	NPS	NPA	100.0% (286/286)	98.7-100%
Seasonal H1	NPW	PPA	Not Detected (0/0)	-
(Clinical Study)	NPW	NPA	99.8% (482/483)	98.9-100%
	NPS	PPA	Not Detected (0/0)	-
	NPS	NPA	100.0% (312/312)	98.8-100%
Seasonal H1	NPS	PPA	100.0% (29/29)	88.1-100%
(Archived Samples)	NPS	NPA	100.0% (21/21)	83.4-100%
Seasonal H1	NPW	PPA	100% (54/54)	93.4-100%
(Surrogate Samples)	NPW	NPA	100.0% (8/8)	63.1-100%
	NPS	PPA	100% (59/59)	93.9-100%
	NPS	NPA	100.0% (7/7)	59.0-100%

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

Clinical Performance (Prospective Study):
- Sample Size: 795 valid specimens (312 NPS, 483 NPW) were analyzed. 6% (44/795) required retesting due to invalid/inconclusive/unsubtypeable initial results.
- Data Provenance: Prospective, collected from 5 geographically separated military clinical sites in the U.S. during the 2010-2011 influenza season (December 2010 to April 2011).
Testing of Preselected Archived Samples (for Seasonal Influenza A/H1):
- Sample Size: 51 NPS specimens (30 known positive seasonal Influenza A/H1, 21 influenza-negative).
- Data Provenance: Retrospective, pre-selected archived clinical NPS specimens obtained and confirmed at two different clinical study sites in the U.S.
Testing of Surrogate Clinical Specimens (for Seasonal Influenza A/H1):
- Sample Size: 136 individual influenza-negative clinical specimens (68 NPS, 68 NPW) were spiked with known concentrations of seasonal Influenza A/H1 virus. Valid results obtained for 128 (62 NPW, 66 NPS).
- Data Provenance: Contrived clinical samples generated from residual influenza-negative NPS and NPW samples. Sent to two different clinical trial sites in the U.S.

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

The document does not explicitly state the number or qualifications of individual experts who established the ground truth. However, the ground truth for the clinical and archived samples was established using a reference method:

"The reference method was the CDC rRT-PCT Flu Panel influenza A and influenza B assays, followed by subtype-specific PCR and bi-directional sequencing of amplicons."
"the presence of Influenza A/H1 viral RNA was confirmed using 'validation' PCR assays. The validation PCR assays were identical to the comparator assays that were used for the prospective clinical evaluation study."

This implies that the "experts" were the personnel performing and interpreting the CDC assays and sequencing results, which would typically be highly trained laboratory professionals and molecular biologists proficient in these techniques, likely adhering to CDC protocols. No specific number of such experts is given.

4. Adjudication Method for the Test Set

The document states: "JBAIDS Software analyzes the fluorescence amplification curves and reports results as positive, negative, or uncertain. A failure of the Positive or Negative Control will result in the entire run being called invalid. Retesting is required to resolve uncertain or invalid results."

For the clinical study, 6% of specimens (44/795) required retesting. Specifically, "Invalid" (32/44), "Inconclusive" (1/44), and "Unsubtypeable" (1/44). "Forty (40) out of 44 samples resolved upon a 1st retest and the remaining 4 samples required a re-extraction and retest and resolved." This indicates a retesting and re-extraction protocol for initial indeterminate or invalid results rather than a multi-expert adjudication panel for interpretive discrepancies. The ground truth itself (CDC rRT-PCR + sequencing) would have its own internal validation/adjudication processes, but this is not detailed for external review.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No multi-reader, multi-case (MRMC) comparative effectiveness study comparing human readers with and without AI assistance was performed. This device is a diagnostic kit (real-time RT-PCR assay) that provides automated qualitative results, not an imaging AI diagnostic aid for human interpretation. Therefore, the concept of "human readers improve with AI vs without AI assistance" does not directly apply to this type of device.

6. Standalone Performance Study (Algorithm Only Without Human-in-the-Loop Performance)

Yes, the studies described (clinical performance, archived samples, surrogate samples) represent standalone performance of the algorithm/device. The "JBAIDS Software analyzes the fluorescence amplification curves and reports results as positive, negative, or uncertain." While human operators perform the physical tests, the interpretation of the raw PCR data into a final qualitative result (positive/negative/uncertain) is automated by the device's software (algorithm). The clinical performance data presented (PPA and NPA) are a direct measure of this standalone performance against a defined ground truth.

7. Type of Ground Truth Used

The ground truth used was:

Expert Consensus/Reference Method: For prospective clinical specimens and archived samples, the reference method was the CDC rRT-PCR Flu Panel influenza A and influenza B assays, followed by subtype-specific PCR and bi-directional sequencing of amplicons. This is a highly robust and accepted laboratory method for viral identification and subtyping.
Known Spiked Concentration: For surrogate clinical specimens, the ground truth for positive samples was established by spiking influenza-negative clinical samples with known concentrations of seasonal Influenza A/H1 virus. Un-spiked negative samples also served as ground truth negatives.

8. Sample Size for the Training Set

The document does not provide details on a specific "training set" sample size or data for the development of the device's algorithm, as would typically be described for machine learning or AI models. This device is a PCR-based assay, where the "algorithm" is primarily the pre-defined thresholds and analysis logic within the JBAIDS software to interpret PCR amplification curves. Performance characteristics are primarily established through analytical validation (LoD, inclusivity, exclusivity) and clinical validation with independent test sets, rather than an explicit training-validation split in the context of an adaptive or learning algorithm. The design of the primers and probes, and the associated software interpretation, would have been iteratively developed and optimized, but a formal "training set" as understood in AI/ML is not mentioned.

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

As noted above, a formal "training set" for an AI/ML algorithm is not described for this diagnostic kit. The underlying principles of the PCR assay (primer/probe design, reaction conditions) are established through extensive analytical studies to ensure specificity and sensitivity. The "ground truth" for developing the analytical performance characteristics (like LoD, exclusivity, inclusivity) would involve testing well-characterized viral strains and non-target organisms with known concentrations, typically obtained from reference collections. This process establishes the analytical sensitivity and specificity that the device is designed to achieve when interpreting a sample.

Summary

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K 111778

SEP 1 3 2011 510(k) Summary JBAIDS Influenza A Subtyping Kit

According to the requirements of 21 CFR 807.92, the following information Introduction: provides sufficient detail to understand the basis for a determination of substantial equivalence.
Submitted by: U.S. Army Medical Materiel Development Activity Division of Regulated Activities and Compliance 1430 Veterans Drive Fort Detrick, MD
- Robert Miller Ph.D., RAC Primary Contact: Director, Division of Regulated Activities and Compliance Telephone: 301-619-0317 Facsimile: 301-619-0197 e-mail: usamrmcregulatoryaffairs(@amedd.army.mil
- Secondary Contact: Patricia Beverly, RAC Division of Regulated Activities and Compliance Telephone: 301-619-2980 Facsimile: 301-619-0197

e-mail: patricia.m.beverly@us.army.mil or usamrmcregulatoryaffairs@amedd.army.mil

Beth Lingenfelter Technical Contact: Director of Regulatory Affairs, Idaho Technology, Inc. Telephone: 801-736-6354, ext 407 Facsimile: 801-588-0507 e-mail: bethl@idahotech.com
Date Prepared: August, 2011

Device Name: Trade Name:

JBAIDS Influenza A Subtyping Kit

Common Name:

Real-time PCR assay for differentiation of influenza A subtypes

Classification Name:

Reagents for Detection of Specific Novel Influenza A Viruses (CFR 866.3332)

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Intended Use

The Joint Biological Agent Identification and Diagnostic System (JBAIDS) Influenza A Subtyping Kit is intended for the in vitro qualitative detection and differentiation of seasonal Influenza A/H1, seasonal Influenza A/H3, and 2009 H1N1Influenza viral nucleic acids isolated and purified from nasopharyngeal swab (NPS) and nasopharyngeal wash (NPW) specimens from human patients with signs and symptoms of respiratory infection, in conjunction with clinical and epidemiological risk factors. The JBAIDS Influenza A Subtyping Kit contains reverse transcriptase real-time polymerase chain reaction (rRT-PCR) assays for use on the JBAIDS instruments. The Flu A H1, Flu A H3, and Flu A H1 2009 assays of the JBAIDS Influenza A Subtyping Kit target a region of the hemagglutinin (HA) gene of the respective Influenza A virus. The Flu A Sw assay of the JBAIDS Influenza A Subtyping Kit targets a region of the nucleocapsid protein (NP) gene of the 2009 H1N1 Influenza virus, as well as some other Influenza A viruses of swine lineage. This kit is not intended to detect Influenza B or Influenza C viruses.

Due to low seasonal prevalence, performance characteristics for detection of seasonal Influenza A/H1 were established primarily with retrospective and contrived clinical specimens.

Device Description

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influenza A/H1, A/H3, and A/2009 H1 viral RNA. In particular, the Flu A H1, Flu A H3, and Flu A H1 2009 assays target distinct regions of the hemagglutinin gene specific to those subtypes, and the Flu A Sw assay targets a region of the nucleocapsid protein gene as a secondary target for the influenza A/2009 H1(swine lineage) virus. The tests are performed using the JBAIDS instrument and software.

Assay Principle

Before testing, NPS or NPW specimens are purified using 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 along with reconstitution buffer. When viral RNA is present, a fragment of influenza A 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 or Negative Control will result in the entire run being called invalid. Retesting is required to resolve uncertain or invalid results.

Substantial Equivalence

The JBAIDS Influenza A Subtyping System is substantially equivalent to other products in commercial distribution intended for similar use. The JBAIDS instrument has been previously cleared under K051713.

The JBAIDS Influenza A Subtyping System is substantially equivalent to the CDC Human Influenza Virus real-time RT-PCR Detection and Characterization Panel, which was cleared on September 30, 2008 under 510(k) # K080570, and the CDC Influenza 2009 A(H1N1)pdm Real-Time RT-PCR Panel, which was cleared on June 22, 2010 under 510(k) #101564.

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Element	JBAIDS Influenza A Subtyping Kit	CDC rRT-PCR Flu Panel (K080570)	CDC rRT-PCR 2009 A(H1N1)pdm Flu Panel (K101564)
Intended Use	Qualitative in vitro detection and differentiation of seasonal Influenza A/H1, seasonal Influenza A/H3 and Influenza A/2009 H1N1 viral nucleic acids from nasopharyngeal swab (NPS) and nasopharyngeal wash (NPW) specimens on the JBAIDS instrument after obtaining an influenza A positive test from the JBAIDS Influenza A & B Detection Kit.	Qualitative in vitro detection of influenza virus type A or B and for determination of the subtype of seasonal human influenza A virus, as seasonal A/HI or A/H3, if present, from viral RNA in nasopharyngeal and/or nasal swab specimens, for presumptive identification of virus in patients who may be infected with influenza A subtype A/H5 (Asian lineage) from viral RNA in human respiratory specimens and viral culture on an ABI 7500 Fast Dx Real-time PCR instrument	Qualitative in vitro detection of influenza virus type A and 2009(H1N1 influenza viral RNA from nasopharyngeal swabs (NPS), nasal swabs (NS), throat swabs (TS), nasal aspirates (NA), nasal washes (NW), dual nasopharyngeal / throat swabs (NPS/TS) and lower respiratory tract specimens (LRTS) from human patients with signs and symptoms of respiratory infection and/or from viral culture, on the Applied Biosystems (ABI) 7500 Fast Dx Real-Time PCR instrument.
Technology	Real-time PCR using hydrolysis probes	Real-time PCR using hydrolysis probes	Real-time PCR using hydrolysis probes
Assay Results	Qualitative	Qualitative	Qualitative
Nucleic Acid Extraction	Yes	Yes	Yes
Table 2. Differences Between the JBAIDS Influenza A Subtyping Kit and its Predicate Devices
Element	JBAIDS Influenza ASubtyping Kit	CDC rRT-PCR Flu Panel(K080570)	CDC rRT-PCR 2009A(H1N1)pdm Flu Panel(K101564)
Viruses Detected	Influenza A/H1, Influenza A/H3and Influenza A/2009 H1N1	Influenza A, Influenza B,Influenza A/H1, Influenza A/H3and Influenza A/H5	Influenza A and Influenza A2009 H1N1
Specimen types	Nasopharyngeal swabs,nasopharyngeal washes	Nasopharyngeal swabs, nasalswabs, and virus culture	Nasopharyngeal swabs, nasalswabs, nasal aspirates, nasalwashes, dual nasopharyngeal /throat swabs, broncheoalveolarlavage, tracheal aspirate,bronchial wash and viral culture
RequiredInstrumentation	JBAIDS instrument	Applied Biosystems 7500 FastDx Real-time PCR instrumentwith SDS software v 1.4	Applied Biosystems 7500 FastDx Real-time PCR instrument
Interpretation ofTest Results	Automated analysis of testresults and controls	User required to interpret testand control results	User required to interpret testand control results
Enzyme MasterMix	Assays come in freeze-driedsingle use vials that include allcomponents of master mix	Invitrogen SuperScript™ IIIPlatinum® One-StepQuantitative RT-PCR Kits	Invitrogen SuperScript™ IIIPlatinum® One-StepQuantitative RT-PCR Kits
Reagent Storage	Reagents are stored at roomtemperature	Reagents are stored at ≤ -20°C	Reagents are stored at ≤ -20°C
ExtractionMethods	• IT 1 - 2 -3™ Platinum PathSample Purification Kit	• Qiagen QIAamp® Viral RNAMini Kit	• Qiagen QIAamp® Viral RNAMini Kit
	• Roche MagNA Pure CompactNucleic Acid Isolation Kit I	• Qiagen RNeasy® Mini Kit• Roche MagNA Pure TNA Kit	• Roche MagNA Pure CompactNucleic Acid Isolation Kit IRoche MagNA Pure TNA Kit
		• Roche MagNA Pure LC RNAIsolation Kit II	• Roche MagNA Pure LC RNAisolation Kit II
			• Qiagen QIAcube withQIAamp viral RNA mini kit• bioMerieux NucliSENSeasyMAG

Table 1. Similarities Between the JBADS Influenza A Subtyping Kit and its Predicate Devices

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2. Differences Between the IRAIDS Influenza A Subtyning Kit and its Predicate Devices

Summary of Performance Data

Clinical Performance

The clinical performance of the JBAIDS Influenza A Subtyping Kit was evaluated during a prospective study at 5 geographically separated military clinical sites over the 2010-2011 influenza season (December 2010 to April 2011). Subjects with signs and/or symptoms of influenza-like illness were enrolled. Upon obtaining informed consent,

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NPS and NPW specimens were collected for JBAIDS and comparator testing. A total of 795 valid specimens were analyzed at the five study sites: 312 NPS and 483 NPW specimens. Table 3 provides a summary of demographic information for the 795 subjects for which valid specimen results were obtained in the prospective study.

		Overall	Site 1	Site 2	Site 3	Site 4	Site 5
NPS		312	રે રેપ	206	રેર	0	0
NPW		483	320	0	0	118	વર્ષ રે
	Total	795	370	206ર્સ		118	45
	Female	405 (50.9%)	188 (50.8%)	122 (59.2%)	23 (41.1%)	56 (47.5%)	16 (35.6%)
Sex	Male	390 (49.1%)	182 (49.2%)	84 (40.8%)	33 (58.9%)	62 (52.5%)	29 (64.4%)
	Mean	26.4	23.3	24.5	30.3	23.1	30.8
	Median	24.0	24.0	18.0	27.5	17.0	26.0
Ageª	Min	0.5	0.5	0.5	2.0	0.5	18.0
	Max	92.0	92.0	69.0	81.0	68.0	62.0
	્રર	149 (18.7%)	88 (23.8%)	40 (19.4%)	4 (7.1%)	17 (14.4%)	0 (0%)
AgeRange®	6-21°	229 (28.8%)	79 (21.4%)	74 (35.9%)	10 (17.9%)	54 (45.8%)	12° (26.7%)
	22-49	331 (41.6%)	178 (48.1%)	54 (26.2%)	35 (62.5%)	34 (28.8%)	30 (66.7%)
	>50	86 (10.8%)	25 (6.8%)	38 (18.4%)	7 (12.5%)	13 (11%)	3 (6.7%)

Table 3. Demographic Summary for the JBAIDS Influenza A Subtyping Kit Prospective Study.

4 0.5 was used for all ages under 1 year for these calculations.

b The age groups ≤ 5 years and ≥ 50 years correspond to high risk groups for which the CDC strongly recommends seasonal influenza vaccination (http://www.cdc.goy/flu//flu/protect/keyfacts.htm). 6 Site 5 enrolled adults only; this category reflects participants 18 to 21 years of age

Of the 795 prospective specimens, successful results were obtained for 94% (751/795) of these specimens on the first attempt (Site 1: 359/370 =97%; Site 2: 191/206 =93%; Site 3: 54/56 =96%; Site 4: 110/118 =93%; Site 5: 37/45 =98%). The remaining 6% (44/795) required retesting: "Invalid" (32/44), "Inconclusive" (1/44), and "Unsubtypeable" (1/44) (11 samples from Site 1; 15 samples from Site 2; 2 samples from Site 3; 8 sample from Site 4: and 8 sample from Site 5). Forty (40) out of 44 samples resolved upon a 1st retest and the remaining 4 samples required a re-extraction and retest and resolved.

Nucleic acid from each specimen was isolated using either the IT I-2-3 Platinum Path Sample Purification Kit (manual sample processing) or the Roche MagNA Pure Compact Nucleic Acid Isolation Kit I (automated sample processing) and tested with the JBAIDS Influenza A Subtyping Kit. The performance of the JBAIDS Influenza A Subtyping Kit was evaluated by comparing the JBAIDS test results with a comparator/reference method. The reference method was the CDC rRT-PCT Flu Panel influenza A and influenza B assays, followed by subtype-specific PCR and bi-directional sequencing of amplicons. Clinical sensitivity was calculated as Positive Percent Agreement (PPA) and specificity was calculated as Negative Percent Agreement (NPA). The exact binomial

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two-sided 95% confidence interval was calculated. The results are summarized in Table 4.

Influenza AStrain	SampleMatrix	PurificationKit	PPA			NPA
			TP/(TP+FN)	Percent	95% CI	TN/(TN+FP)	Percent	95% CI
	Seasonal H1	NPW	Platinum Path	0/0	-	-	277/278	99.6%
	NPW	MagNA Pure	0/0	-	-	205/205	100.0%	98.2-100%
		Combined	0/0	-	-	482/483	99.8%	98.9-100%
	NPS	Platinum Path	0/0	-	-	132/132	100.0%	97.2-100%
	NPS	MagNA Pure	0/0	-	-	180/180	100.0%	98.0-100%
		Combined	0/0	-	-	312/312	100.0%	98.8-100%
	NPW	Platinum Path	50/50	100.0%	92.9-100%	227/228	99.6%	97.6-100%
2009 H1N1		MagNA Pure	16/16	100.0%	79.4-100%	187/189	98.9%	96.2-99.9%
		Combined	66/66	100.0%	94.6-100%	414/417	99.3%	97.9-99.9%
	NPS	Platinum Path	24/24	100.0%	85.8-100%	108/108	100.0%	96.6-100%
		MagNA Pure	10/10	100.0%	69.2-100%	169/170	99.4%	96.8-100%
		Combined	34/34	100.0%	89.7-100%	277/278	99.6%	98.0-100%
	NPW	Platinum Path	14/14	100.0%	76.8-100%	264/264	100.0%	98.6-100%
Seasonal H3		MagNA Pure	19/19	100.0%	82.4-100%	186/186	100.0%	98.0-100%
		Combined	33/33	100.0%	89.4-100%	450/450	100.0%	99.2-100%
	NPS	Platinum Path	18/18	100.0%	81.5-100%	115/115	100.0%	96.8-100%
		MagNA Pure	8/8	100.0%	63.1-100%	171/171	100.0%	97.9-100%
		Combined	26/26	100.0%	86.8-100%	286/286	100.0%	98.7-100%

Table 4. JBAIDS Influenza A Subtyping Kit Prospective Clinical Performance Summary

Seasonal influenza A/H1 virus was not circulating during the 2010-2011 influenza season (http://www.cdc.gov/flu/) and was not detected during the prospective clinical study of the JBAIDS Influenza A Subtyping Kit. To supplement the results of the clinical study, an evaluation of preselected archived samples was performed. Due to the limited availability of archived specimens, the clinical study was further supplemented with surrogate clinical contrived specimens.

Testing of Preselected Archived Specimens

Additional testing of pre-selected archived clinical NPS specimens was performed at two different clinical study sites to supplement the prospective clinical testing data. Because it is possible that the archived samples had been misidentified or had degraded during storage or previous handling, the presence of Influenza A/H1 viral RNA was confirmed using "validation" PCR assays. The validation PCR assays were identical to the comparator assays that were used for the prospective clinical evaluation study. A total of 51 NPS specimens were obtained and confirmed for testing: 30 known to be positive seasonal Influenza A/H1 specimens and 21 influenza-negative specimens. Validated samples were purified using either the IT 1-2-3 Platinum Path Sample Purification Kit or

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the Roche MagNA Pure Compact Nucleic Acid Isolation Kit I and then tested with the Flu A and human sample control (Flu SC) assay from the JBAIDS Influenza A & B Detection Kit and the Flu A H1 assay from the JBAIDS Influenza A Subtyping Kit. The specimens were split evenly for purification with the Platinum Path or MagNA Pure purification kits and then randomized such that the users performing the JBAIDS testing were blinded as to the expected test result.

Table 5 presents the PPA and NPA for the archived clinical specimens. Data from both extraction kits are combined due to identical performance.

InfluenzaAssay	SampleType	PPA	Percent	95% CI	NPA	Percent	95% CI
Flu A H1	NPS	29/29	100%	88.1-100%	21/21	100%	83.4-100%

Table 5, Performance Summary of Seasonal Influenza A/H1 Archived Clinical Specimens

Due to the absence of seasonal Influenza A/H1 virus in circulation during the 2010-2011 influenza seasonal (http://www.cdc.gov/flu/) and lack of availability of archived NPW specimens for seasonal Influenza A/H1, contrived clinical samples (residual influenza negative NPS and NPW samples spiked with a known concentration of seasonal Influenza A/H1 virus) were used as a surrogate to further evaluate the performance of the JBAIDS Influenza A Subtyping Kit.

Testing of Surrogate Clinical Specimens

A total of 136 individual influenza-negative clinical specimens (68 NPS samples and 68 NPW samples) were spiked at a range of concentrations, including near the system limit of detection (LoD), as well as un-spiked, then randomized, and sent to two different clinical trial sites for testing. Of the 136 surrogate samples included in this study, a valid JBAIDS test result was obtained for 128 samples (62 NPW and 66 NPS). The remaining 8 samples with invalid results could not be retested due to insufficient sample volume, and were not included in the analysis.

Table 6 presents the PPA and NPA for the surrogate clinical specimens. Half of the samples were extracted using the Platinum Path purification kit and half using the MagNA pure kit. Performance from both extraction kits was identical, so results are combined.

Assay	Sample Type	PPA			NPA
		TP/(TP+FN)	Percent	95% CI	TN/(TN+FP)	Percent	95% CI
Flu A H1	NPW	54/54	100%	93.4-100%	8/8	100.0%	63.1-100%
Flu A H1	NPS	59/59	100%	93.9-100%	7/7	100.0%	59.0-100%
Flu A H1 2009/Flu A Sw	NPW	0/0	-	-	62/62	100.0%	94.2-100%
Flu A H1 2009/Flu A Sw	NPS	0/0	-	-	66/66	100.0%	94.6-100%
Flu A H3	NPW	0/0	-	-	62/62	100.0%	94.2-100%
Flu A H3	NPS	0/0	-	-	66/66	100.0%	94.6-100%

Table 6. Performance Summary of Seasonal Influenza A/H1 Surrogate Clinical Specimens

U.S. Army Office of the Surgeon General. 510(k) June, 2011 JBAIDS Influenza A Subtyping System

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Analyses of the clinical data set, preselected archived specimens, and surrogate clinical specimens demonstrate that the JBAIDS Influenza A Subtyping Kit is a sensitive and specific test system for the differentiation of influenza A/H1, influenza A/H3, and influenza A/2009 H1 virus subtypes.

Selected Analytic Studies

Limit of Detection

The analytical sensitivity or Limit of Detection (LoD) for each target assay (Flu A H1, Flu A H3, Flu A H1 2009, and Flu A Sw) was determined using both NPS and NPW samples spiked with quantified virus strains. The LoD was the lowest concentration where ≥ 95% of samples yielded positive results. Twenty (20) independent specimens from 20 unique donors were spiked with each virus strain for each sample type/purification kit combination and tested at the LoD concentration. The LoD values for representative virus strains detected by the JBAIDS Influenza A Subtyping Kit are listed in Table 7.

Subtyping Kit
Assay(s)	Influenza Type	Strain	LoD(EID50/mL)
Flu A H1	Influenza A H1N1	A/New Caledonia/20/1999	50a
		A/Hawaii/15/2001	5,000
Flu A H3	Influenza A H3N2	A/New York/55/2004	5
		A/Wisconsin/67/2005	10
Flu A H1 2009and Flu A Sw	2009 H1N1 Influenza	A/New York/18/2009	1,500
		A/California/7/2009	5,000

Table 7. LoD Concentrations for Representative Virus Strains Detected by the JBAIDS Influenza A
Subtyping Kit

4 The aliquot of A/New Caledonia/20/1999 tested contains 17-45 times more PCR target copies per E/Dg than the aliquot of A/Hawaii/15/2001 tested.

Inclusivity

The analytical reactivity of the JBAIDS Influenza A Subtyping Kit assays was evaluated with inclusivity panels consisting of eight seasonal influenza A H1N1 strains (Table 8), 10 seasonal influenza A H3N2 strains (Table 9), and 11 2009 H1N1 influenza strains (Table 10) that represent the genetic, temporal, and geographic diversity of the influenza analytes. Each organism was tested in a simulated NPS sample matrix at or near the system LoD (5, 50, and 500 EID50/mL or TCID50/mL for H1N1 strains; 0.5, 5 and 50 EID50/mL or TCID50/mL for H3N2 strains; and 150, 1,500 and 15,000 EID50/mL or TCIDsofmL for H1N1 2009 strains). Higher concentrations were tested if the analyte was not detected at the initial test concentrations. Four (4) of the 29 influenza strains tested in this study were not detected with the appropriate JBAIDS influenza A subtyping assays.

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There was considerable variability in the ability of the Flu A H1 assay to detect strains (lowest detected concentrations ranged from 5-500,000 TCID30/mL). Sequence alignments of tested strains indicate variability potentially due to mismatches under the primers and probe. The influenza A/1/Denver/1/57 strain was not detected at a final concentration of 5,000 TCID50/mL.

For the Flu A H3 assay, the following strains were not detected at the following concentration: A/Aichi/2/68 at 114,000 TCID50/mL, A/Hong Kong/8/68 at 137,000 TCID50/mL, and A/MRC-2 recomb at 7,350 TCID30/mL. Sequence alignments indicate that the A/Aichi/2/68 and A/Hong Kong/8/68 isolates have mismatches in the probe sequence. The sequence for A/MRC-2 recomb was not available.

For the Flu A H1 and Flu A H3 assays, in silico evaluation of contemporary strains (2006-2011) indicate that there are few mismatches, and strains should be detected.

Strain	LowestConcentrationDetected
A/PR/8/34	500,000 TCID50/mL
A/NWS/33	50 TCID50/mL
A/Weiss/43	500 TCID50/mL
A1/FM/1/47	5 TCID50/mL
A/Mal/302/54	5,000 TCID50/mL
A1/Denver/1/57	NDª
A/Solomon Islands/3/2006	5 TCID50/mL
A/Brisbane/59/07	50 TCID50/mL

	Table 8. Results of Influenza A H1N1 Inclusivity

8 ND stands for 'not detected'

Strain	Lowest ConcentrationDetected
A/Aichi/2/68	NDa
A/Hong Kong/8/68	NDa
A/Port Chalmers/1/73	5 TCID50/mL
A/Victoria/3/75	50 TCID50/mL
A/Brisbane/10/07	5 TCID50/mL
A/Taiwan/760/2007	0.5 TCID50/mL
A/Uruguay/716/2007	0.5 EID50/mL
A/Perth/16/09	5 TCID50/mL
A/Alice	0.5 TCID50/mL
A/MRC-2 recomb	NDa

Table 9. Results of Influenza A H3N2 Inclusivity

4 ND stands for 'not detected'

June, 2011

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Strain	Lowest ConcentrationDetected
A/California/4/2009	1,500 TCID50/mL
A/California/8/2009	150 EID50/mL
A/England/195/2009	150 TCID50/mL
A/Mexico/4108/2009	150 EID50/mL
A/North Carolina/18/2009	1,500 TCID50/mL
A/South Carolina/18/2009	150 TCID50/mL
A/SwineNY/01/2009	150 TCID50/mL
A/SwineNY/02/2009	150 TCID50/mL
A/SwineNY/03/2009	150 TCID50/mL
A/Texas/48/2009	1,500 TCID50/mL
A/Washington/29/2009	150 TCID50/mL

Table 10. Results of 2009 H1N1 Influenza Inclusivity

Exclusivity

The potential for cross-reactivity between JBAIDS influenza assays was evaluated by testing simulated NPS samples containing high concentrations of influenza viruses (tens to thousands-fold higher than LoD). Table 11 lists all of the non-target influenza strains tested at high concentrations with the Flu A H1, Flu A H3, Flu A Sw and Flu A H1 2009 assays. In all cases, the assays gave the expected negative results with the non-target influenza assays.

Three (3) Influenza A H1N1 strains, A/Maryland/12/1991, A/Iowa/1/2006, and A/swine/Wisconsin/125/1997, were detected by the Flu A Sw assay: These results were not unexpected since the first two of these strains were isolated from humans but have an origin of swine lineage and the third was isolated from swine. In addition, the Influenza A H3N2 virus strain A/SW/IA/1/99 (swine origin) was detected by both the Flu A H3 and Flu A Sw assays. Detection of swine Influenza A/H3 viruses by the Flu A H3 assay is not unexpected as the hemagglutinin sequences for swine and human isolates are very similar.

	Type/Subtype	Strain	ConcentrationTested	AssaysTested
Influenza A	H2N2 (Avian)	A/chicken/Pennsylvania/298101-4/2004	3.16E+07 TCID50/mL	Flu A H1
	H3N8 (Avian)	A/MAL/ALB/16/87	1.72E+03 TCID50/mL	Flu A H3
	H4N8 (Avian)	A/chicken/Alabama/1975	1.00E+08 EID50/mL	Flu A Sw
	H5N1 (Avian-HumanRecombinant)	A/Vietnam/1203/2004(H5N1)-PR8	3.16E+07 EID50/mL	Flu A H12009
	H5N1 (Avian)	A/DK/PA/4560069-9/06	1.00E+05 TCID50/mL
	H7N3 (Avian)	A/TY/UT/24721-10/95	3.06E+04 TCID50/mL

Table 11. Results of Testing for Cross-Reactivity with Influenza A Subtyping Assay
Virus	Subtype	Assay Result	Virus	Subtype	Assay Result
Adenovirus	Type 1	Negative	Influenza B	Victoria Lineage	Negative
Adenovirus	Type 3	Negative	Influenza B	Yamagata Lineage	Negative
Coronavirus	229E	Negative	Metapneumovirus		Negative
Coronavirus	HKU1	Negative	Parainfluenza	Type 1	Negative
Coronavirus	NL63	Negative	Parainfluenza	Type 2	Negative
Coronavirus	OC43	Negative	Parainfluenza	Type 3	Negative
Enterovirus	EV-D68	Negative	Rhinovirus		Negative
Enterovirus		Negative	RSV		Negative

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Type/Subtype	Strain	ConcentrationTested	AssaysTested
H6N2 (Avian)	A/Chicken/CA/32213-1/2000	1.26E+07 EID50/mL
H9N2 (Avian)	A/Turkey/Wisconsin/1966	5.60E+07 EID50/mL
H3N8 (Canine)	A/canine/Florida/43/2004	1.00E+05 TCID50/mL
H3N8 (Equine)	A/Equine/Ohio/01/2009	1.00E+05 TCID50/mL
H1N1 (Swine)	A/swine/Wisconsin/125/1997	1.00E+05 TCID50/mL
H1N1 (Swine)	A/SW/GB/19582/92	5.64E+03 TCID50/mL
H3N2 (Swine)	A/SW/IA/1/99	1.41E+03 TCID50/mL
H1N1 (Human ofswine lineage)	A/Maryland/12/1991	1.00E+05 TCID50/mL
H1N1 (Human ofswine lineage)	A/Iowa/1/2006	1.00E+05 TCID50/mL
H7N2 (Human)	A/New York/107/2003	30 µl of an unknownconcentration into1mL
	B/Lee/40	7.36E+03 TCID50/mL
	B/Allen/45	1.00E+05 TCID50/mL
	B/GL/1739/54	7.36E+03 TCID50/mL	Flu A H1
	B/Maryland/1/59	7.36E+03 TCID50/mL	Flu A H3
Influenza B	B/Taiwan/2/62	4.54E+04 TCID50/mL	Flu A Sw
	B/Hong Kong/5/72	7.36E+03 TCID50/mL	Flu A H12009
	B/Malaysia/2506/04	5.09E+03 TCID50/mL
	B/FL/04/06	1.50E+04 TCID50/mL
	B/Brigit	3.14E+04 TCID50/mL
	A/Brisbane/59/07	1.00E+05 TCID50/mL
	A1/FM/1/47	4.24E+03 TCID50/mL
	A/PR/8/34	1.00E+05 TCID50/mL	Flu A H3
Influenza AH1N1	A/NWS/33	4.24E+03 TCID50/mL	Flu A Sw
	A1/Denver/1/57	4.24E+03 TCID50/mL	Flu A H12009
	A/Solomon Islands/3/2006	1.25E+04 TCID50/mL
	A/Weiss/43	4.24E+03 TCID50/mL
	A/Mal/302/54	1.25E+04 TCID50/mL
	A/Port Chalmers/1/73	5.10E+03 TCID50/mL
	A/Victoria/3/75	4.24E+03 TCID50/mL
	A/Aichi/2/68	1.00E+05 TCID50/mL	Flu A H1
Influenza AH3N2	A/Hong Kong/8/68	1.00E+05 TCID50/mL	Flu A SwFlu A H12009
	A/Alice (VR-776)	4.24E+03 TCID50/mL
	A/MRC-2 recomb (VR-777)	7.36E+03 TCID50/mL
	A/Brisbane/10/07	7.36E+03 TCID50/mL
Influenza A(swine lineage)H1N1 2009	Swine NY/02/2009	1.25E+04 TCID50/mL	Flu A H1
	Swine NY/03/2009	7.36E+03 TCID50/mL	Flu A H3
	Swine NY/01/2009	3.78E+04 TCID50/mL
	A/Mexico/4108/2009	1.00E+05 EID50/mL
	A/California/8/2009	1.00E+05 TCID50/mL
	A/California/04/2009	1.00E+05 TCID50/mL
Type/Subtype	Strain	ConcentrationTested	AssaysTested
	A/Washington/29/2009	1.00E+05 TCID50/mL
	A/South Carolina/18/2009	1.00E+05 TCID50/mL
	A/England/195/2009	4.74E+04 TCID50/mL
	A/North Carolina/39/2009	1.00E+05 TCID50/mL

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The non-influenza exclusivity panel consisted of 17 bacteria, 18 viruses, and one fungus, which were selected based on the relatedness to JBAIDS influenza analytes, clinical relevance (cause respiratory symptoms or represent nasopharyngeal flora), or high prevalence within the population (e.g. Herpes Simplex Virus). Simulated NPS samples were spiked with bacteria or fungi at a concentration of 10° CFU/mL or TCID30/mL and viruses at a concentration between 103 - 10' copies/mL or TCID50/mL. The JBAIDS Influenza A subtyping assays did not cross-react with the exclusivity panel organisms at the test concentrations listed in Table 12.

Virus	Concentration Tested	Bacteria/Fungi	Concentration Tested
Adenovirus	1.00E+05 TCID50/mL	Bordetella pertussis	1.00E+06 CFU/mL
Bocavirus	4.20E+07 copies/mL	Candida albicans	1.00E+06 CFU/mL
Coronavirus 229E	7.35E+03 TCID50/mL	Corynebacterium diptheriae	1.00E+06 CFU/mL
Coronavirus OC43	6.57E+04 TCID50/mL	Escherichia coli	1.00E+06 CFU/mL
Coronavirus NL63	5.10E+03 TCID50/mL	Haemophilus influenza	7.80E+04 CFU/mL
Coronavirus HKU1	1.00E+05 copies/mL	Lactobacillus plantarum	1.00E+06 CFU/mL
Cytomegalovirus (CMV)	1.50E+04 TCID50/mL	Legionella pneumophila	1.00E+06 TCID50/mL
Enterovirus	1.00E+05 TCID50/mL	Moraxella catarrhalis	1.00E+06 CFU/mL
Epstein-Barr Virus (EBV)	1.00E+05 copies/mL	Mycobacterium tuberculosis	1.00E+06 CFU/mL
Human Metapneumovirus	7.35E+03 TCID50/mL	Mycoplasma pneumonia	1.69E+05 TCID50/mL
Human Rhinovirus	5.10E+03 TCID50/mL	Neisseria elongata	1.00E+06 CFU/mL
Measles Virus (Rubeola)	1.00E+05 TCID50/mL	Neisseria meningitidis	1.00E+06 CFU/mL
Mumps	4.53E+04 TCID50/mL	Pseudomonas aeruginosa	1.00E+06 CFU/mL
Parainfluenza virus 1	1.25E+04 TCID50/mL	Staphylococcus aureus	1.00E+06 CFU/mL
Parainfluenza virus 2	1.50E+04 TCID50/mL	Staphylococcus epidermidis	1.00E+06 CFU/mL
Parainfluenza virus 3	1.00E+05 TCID50/mL	Streptococcus pneumonia	1.00E+06 CFU/mL
Parainfluenza virus 4	1.00E+05 TCID50/mL	Streptococcus pyogenes	1.00E+06 CFU/mL
Respiratory Syncytial Virus	1.25E+04 TCID50/mL	Streptococcus salivarius.	7.59E+05 CFU/mL

Table 12. Non-Influenza Exclusivity Panel

Reproducibility

A multicenter study was performed to determine overall system reproducibility. Reproducibility testing occurred at three test sites utilizing six total panels of NPS and NPW samples, each, were spiked with a representative seasonal influenza A H1N1 virus (A/New Caledonia/20/1999). Panels of simulated NPS and simulated NPW samples, each, were spiked with a representative seasonal influenza A H3N2 virus (A/New York/55/2004). Finally, panels of simulated NPS and simulated NPW samples,

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each, were spiked with a representative 2009 H1N1 influenza virus (A/New York/18/2009). Samples in each panel consisted of three samples spiked below LoD (high negative, LoD/20), three samples spiked with a low concentration of virus (low positive, LoD), and three samples spiked at a medium concentration of virus (medium positive, 3×LoD) for a total of nine samples per panel. Each panel was tested twice daily at each site for a total of 30 results per sample and 90 results per spike level. The detection rate was ≥ 98% for samples containing influenza virus ≥ LoD. As expected, samples spiked below LoD have variable results. Results are shown in Table 13, Table 14 and Table 15.

SampleType	VirusSpikeLevel	IT 1-2-3 Platinum PathSample Purification Kit			Roche MagNA Pure CompactNucleic Acid Isolation Kit I				Both Kits,All Sites(% Pos.)	95% CI
		Site 1	Site 2	Site 3	Overallfor AllSites	Site 1	Site 2	Site 3	Overallfor AllSites
NPS	3xLoD	15/15(100%)	14/15(93%)	15/15(100%)	44/45(98%)	15/15(100%)	14/15(93%)	15/15(100%)	44/45(98%)	88/90(98%)	92.2-99.7
	LoD	15/15(100%)	15/15(100%)	15/15(100%)	45/45(100%)	15/15(100%)	15/15(100%)	15/15(100%)	45/45(100%)	90/90(100%)	96.7-99.9
	Detection≥ LoD	30/30(100%)	29/30(97%)	30/30(100%)	89/90(99%)	30/30(100%)	29/30(97%)	30/30(100%)	89/90(99%)	178/180(99%)	96.0-99.9
	LoD/20	15/15(100%)	13/15(87%)	8/15(53%)	36/45(80%)	13/15(87%)	11/15(73%)	14/15(93%)	38/45(84%)	74/90(82%)	72.7-89.4
	Detectionall Levels	45/45(100%)	42/45(93%)	38/45(84%)	125/135(93%)	43/45(96%)	40/45(89%)	44/45(98%)	127/135(94%)	252/270(93%)	90.0-96.0
NPW	3xLoD	15/15(100%)	15/15(100%)	15/15(100%)	45/45(100%)	15/15(100%)	15/15(100%)	15/15(100%)	45/45(100%)	90/90(100%)	96.7-99.9
	LoD	14/15(93%)	15/15(100%)	15/15(100%)	44/45(98%)	15/15(100%)	14/15(93%)	15/15(100%)	44/45(98%)	88/90(98%)	92.2-99.7
	Detection≥ LoD	29/30(97%)	30/30(100%)	30/30(100%)	89/90(99%)	30/30(100%)	29/30(97%)	30/30(100%)	89/90(99%)	178/180(99%)	96.0-99.9
	LoD/20	13/15(87%)	15/15(100%)	9/15(60%)	37/45(82%)	14/15(93%)	14/15(93%)	15/15(100%)	43/45(96%)	80/90(89%)	80.5-94.5
	Detectionall Levels	42/45(93%)	45/45(100%)	39/45(87%)	126/135(93%)	44/45(98%)	43/45(96%)	45/45(100%)	132/135(98%)	258/270(96%)	92.4-97.7

Table 13. Summary of Reproducibility Testing for the Flu A H1 Assay (Agreement with Expected Positive Results)

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		IT 1-2-3 Platinum PathSample Purification Kit				Roche MagNA Pure CompactNucleic Acid Isolation Kit I
SampleType	VirusSpikeLevel	Number Positive Samples/Total Samples(% Positive Detection)				Number Positive Samples/Total Samples(% Positive Detection)				Both Kits,All Sites(% Pos.)	95% CI
		Site 1	Site 2	Site 3	OverallFor AllSites	Site 1	Site 2	Site 3	OverallFor AllSites
sNPS	3×LoD	15/15(100%)	15/15(100%)	15/15(100%)	45/45(100%)	15/15(100%)	15/15(100%)	13/15(87%)	43/45(96%)	88/90(98%)	92.2-99.7
	LoD	15/15(100%)	15/15(100%)	15/15(100%)	45/45(100%)	15/15(100%)	15/15(100%)	15/15(100%)	45/45(100%)	90/90(100%)	96.7-99.9
	Detection≥ LoD	30/30(100%)	30/30(100%)	30/30(100%)	90/90(100%)	30/30(100%)	30/30(100%)	28/30(93%)	88/90(98%)	178/180(99%)	96.0-99.9
	LoD/20	12/15(80%)	10/15(67%)	9/15(60%)	31/45(69%)	14/15(93%)	13/15(87%)	14/15(93%)	41/45(91%)	72/90(80%)	70.2-87.7
	Detectionall Levels	42/45(93%)	40/45(89%)	39/45(87%)	121/135(90%)	44/45(98%)	43/45(96%)	42/45(93%)	129/135(96%)	250/270(93%)	88.8-95.4
sNPW	3×LoD	15/15(100%)	15/15(100%)	14/15(93%)	44/45(98%)	15/15(100%)	15/15(100%)	15/15(100%)	45/45(100%)	89/90(99%)	94.0-99.9
	LoD	14/15(93%)	15/15(100%)	15/15(100%)	44/45(98%)	15/15(100%)	15/15(100%)	15/15(100%)	45/45(100%)	89/90(99%)	94.0-99.9
	Detection≥ LoD	29/30(97%)	30/30(100%)	29/30(97%)	88/90(98%)	30/30(100%)	30/30(100%)	30/30(100%)	90/90(100%)	178/180(99%)	96.0-99.9
	LoD/20	11/15(73%)	10/15(67%)	9/15(60%)	30/45(67%)	15/15(100%)	14/15(93%)	15/15(100%)	44/45(98%)	74/90(82%)	72.7-89.5
	Detectionall Levels	40/45(89%)	40/45(89%)	38/45(84%)	118/135(87%)	45/45(100%)	44/45(98%)	45/45(100%)	134/135(99%)	252/270(93%)	89.7-96.0

Table 14. Summary of Reproducibility Testing for the Flu A H3 Assay (Agreement with Expected Positive Results)

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Sample Type	Virus Spike Level	IT 1-2-3 Platinum Path Sample Purification Kit				Roche MagNA Pure Compact Nucleic Acid Isolation Kit I				Both Kits, All Sites	95% CI
		Number Positive Samples/ Total Samples (% Positive Detection)				Number Positive Samples/ Total Samples (% Positive Detection)
		Site 1	Site 2	Site 3	Overall for All Sites	Site 1	Site 2	Site 3	Overall for All Sites
sNPS	3×LoD	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	90/90 (100%)	96.7-99.9
	LoD	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	90/90 (100%)	96.7-99.9
	Detection ≥ LoD	30/30 (100%)	30/30 (100%)	30/30 (100%)	90/90 (100%)	30/30 (100%)	30/30 (100%)	30/30 (100%)	90/90 (100%)	180/180 (100%)	98.3-99.9
	LoD/20	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	90/90 (100%)	96.7-99.9
	Detection all Levels	45/45 (100%)	45/45 (100%)	45/45 (100%)	135/135 (100%)	45/45 (100%)	45/45 (100%)	45/45 (100%)	135/135 (100%)	270/270 (100%)	98.9-99.9
sNPW	3×LoD	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	90/90 (100%)	96.7-99.9
	LoD	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	90/90 (100%)	96.7-99.9
	Detection ≥ LoD	30/30 (100%)	30/30 (100%)	30/30 (100%)	90/90 (100%)	30/30 (100%)	30/30 (100%)	30/30 (100%)	90/90 (100%)	180/180 (100%)	98.3-99.9
	LoD/20	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	15/15 (100%)	15/15 (100%)	15/15 (100%)	45/45 (100%)	90/90 (100%)	96.7-99.9
	Detection all Levels	45/45 (100%)	45/45 (100%)	45/45 (100%)	135/135 (100%)	45/45 (100%)	45/45 (100%)	45/45 (100%)	135/135 (100%)	270/270 (100%)	98.9-99.9

Table 15. Summary of Reproducibility Testing for the Combined Results of the Flu A H1 2009 and Flu A Sw Assays (Agreement with Expected Positive Results)

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Image /page/16/Picture/12 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized eagle or bird-like figure with flowing lines, positioned to the right. Encircling the bird is text that reads "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" in a circular arrangement.

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

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

SEP 1 3 201

Re: K111778

Trade/Device Name: JBAIDS Influenza A Subtyping Kit Regulation Number: 21 CFR $866.3332 Reagents for Detection of Specific Novel Influenza A Viruses Regulation Name: Regulatory Class: Class II Product Codes: OOW, OEP, OOL IDated: June 20. 2011 Received: June 23. 2011

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 10 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.

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 Part 801); 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.

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If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please go to http://www.fda.gov/AboutFDA/CentersOffices/CDRH/CDRHOffices/ucm11.5809.htm for the Center for Devices and Radiological Health's (CDRH's) Office of Compliance. 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/Safeiy/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/MedicalDevices/Resourcesfor You/Industry/default.htm.

.
.

Sincerely yours.

Uwe Scharf for

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

Enclosure

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Indications for Use

510(k) Number: K111778 Device Name: JBAIDS Influenza A Subtyping Kit

Indications for Use:

Due to low seasonal prevalence, performance characteristics for detection of seasonal Influenza A/H1 were established primarily with retrospective and contrived clinical specimens.

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Prescription Use X (Part 21 CFR 801 Subpart D)

AND/OR

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

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

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

Tamar Feldsays
Division Sign Off

Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) // 778

Regulation Number and Section

§ 866.3332 Reagents for detection of specific novel influenza A viruses.

(a)
Identification. Reagents for detection of specific novel influenza A viruses are devices that are intended for use in a nucleic acid amplification test to directly detect specific virus RNA in human respiratory specimens or viral cultures. Detection of specific virus RNA aids in the diagnosis of influenza caused by specific novel influenza A viruses in patients with clinical risk of infection with these viruses, and also aids in the presumptive laboratory identification of specific novel influenza A viruses to provide epidemiological information on influenza. These reagents include primers, probes, and specific influenza A virus controls.(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Reagents for Detection of Specific Novel Influenza A Viruses.” See § 866.1(e) for information on obtaining this document.
(2) The distribution of these devices is limited to laboratories with experienced personnel who have training in standardized molecular testing procedures and expertise in viral diagnosis, and appropriate biosafety equipment and containment.