(82 days)
No
The device description and performance studies focus on real-time RT-PCR technology and standard statistical analysis of sensitivity and specificity, with no mention of AI or ML algorithms for data processing or interpretation.
No.
This device is an in vitro diagnostic (IVD) tool designed for the qualitative detection and differentiation of Influenza A subtypes. It identifies specific viral nucleic acids in patient samples to help diagnose an infection, rather than treating or preventing a disease.
Yes
The device 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" from human patients with signs and symptoms of respiratory infection. This process of identifying specific viral nucleic acids to determine the presence and subtype of influenza is a diagnostic function. The "Intended Use" section explicitly states its use "in conjunction with clinical and epidemiological risk factors" and that "Test results are to be used in conjunction with other clinical and epidemiological information," implying its role in informing a diagnosis.
No
The device is a test kit containing reagents (freeze-dried assays with primer and fluorescent-probe sets) for use with a specific instrument and software, making it a combination product involving both hardware and software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The "Intended Use / Indications for Use" section explicitly states that the 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..." The phrase "in vitro" (meaning "in glass" or "outside the body") is a key indicator of an IVD.
- Device Description: The "Device Description" further clarifies that it's 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..." Again, the term "in vitro" is used.
- Specimen Type: The device analyzes specimens (nasopharyngeal swab and wash) taken from the human body, but the analysis itself is performed outside the body.
- Diagnostic Purpose: The results are intended to be used "in conjunction with other clinical and epidemiological information" to aid in the diagnosis of influenza A infection.
These points align with the definition of an In Vitro Diagnostic device, which is a medical device intended for use in vitro for the examination of specimens derived from the human body solely or principally for the purpose of providing information concerning a physiological or pathological state, or concerning a congenital abnormality, or to monitor therapeutic measures.
N/A
Intended Use / Indications for 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.
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.
Product codes (comma separated list FDA assigned to the subject device)
OOW, OEP, OOL
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.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
nasopharyngeal swab (NPS), nasopharyngeal wash (NPW)
Indicated Patient Age Range
Not Found
Intended User / Care Setting
designated Department of Defense (DoD) laboratories equipped with the JBAIDS instruments.
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
Clinical Performance Test Set:
- Sample size: 795 valid specimens (312 NPS and 483 NPW specimens)
- Data source: Prospective study at 5 geographically separated military clinical sites during the 2010-2011 influenza season (December 2010 to April 2011) from human patients with signs and symptoms of respiratory infection.
- Annotation protocol: 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). 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.
Archived Specimens Test Set:
- Sample size: 51 NPS specimens (30 known positive seasonal Influenza A/H1 specimens and 21 influenza-negative specimens).
- Data source: Preselected archived clinical NPS specimens from two different clinical study sites.
- Annotation protocol: The presence of Influenza A/H1 viral RNA was confirmed using "validation" PCR assays, identical to the comparator assays used for the prospective clinical evaluation study. Validated samples were purified using either the IT 1-2-3 Platinum Path Sample Purification Kit or 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. Specimens were split evenly for purification and randomized, blinding users.
Surrogate Clinical Specimens Test Set:
- Sample size: 136 individual influenza-negative clinical specimens (68 NPS samples and 68 NPW samples). Of these, 128 samples (62 NPW and 66 NPS) yielded valid JBAIDS test results.
- Data source: Residual influenza-negative NPS and NPW samples, spiked with a known concentration of seasonal Influenza A/H1 virus, including near the system limit of detection (LoD), as well as un-spiked.
- Annotation protocol: Samples were randomized and sent to two different clinical trial sites for testing. Half of the samples were extracted using the Platinum Path purification kit and half using the MagNA Pure kit.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Clinical Performance Study:
- Study Type: Clinical Performance / Prospective study.
- Sample Size: 795 valid specimens (312 NPS and 483 NPW).
- Standalone Performance: Successful results for 94% (751/795) on first attempt. 6% (44/795) required retesting; 40/44 resolved on 1st retest, remaining 4 required re-extraction and resolved.
- Key Results:
- For 2009 H1N1 in NPW: PPA 100.0% (66/66) [95% CI: 94.6-100%], NPA 99.3% (414/417) [95% CI: 97.9-99.9%].
- For 2009 H1N1 in NPS: PPA 100.0% (34/34) [95% CI: 89.7-100%], NPA 99.6% (277/278) [95% CI: 98.0-100%].
- For Seasonal H3 in NPW: PPA 100.0% (33/33) [95% CI: 89.4-100%], NPA 100.0% (450/450) [95% CI: 99.2-100%].
- For Seasonal H3 in NPS: PPA 100.0% (26/26) [95% CI: 86.8-100%], NPA 100.0% (286/286) [95% CI: 98.7-100%].
- Seasonal Influenza A/H1 was not circulating and not detected in prospective clinical study.
Testing of Preselected Archived Specimens (Seasonal Influenza A/H1):
- Study Type: Evaluation of Archived Samples.
- Sample Size: 51 NPS specimens (30 known positive, 21 influenza-negative).
- Key Results:
- Flu A H1 for NPS: PPA 100% (29/29) [95% CI: 88.1-100%], NPA 100% (21/21) [95% CI: 83.4-100%].
Testing of Surrogate Clinical Specimens (Seasonal Influenza A/H1):
- Study Type: Evaluation of Contrived Samples.
- Sample Size: 128 valid samples (62 NPW, 66 NPS) out of 136 prepared.
- Key Results:
- Flu A H1 for NPW: PPA 100% (54/54) [95% CI: 93.4-100%], NPA 100.0% (8/8) [95% CI: 63.1-100%].
- Flu A H1 for NPS: PPA 100% (59/59) [95% CI: 93.9-100%], NPA 100.0% (7/7) [95% CI: 59.0-100%].
Limit of Detection (LoD) Study:
- Study Type: Analytical sensitivity study.
- Sample Size: 20 independent specimens from 20 unique donors for each virus strain/sample type/purification kit combination.
- Key Results: LoD for representative virus strains at which ≥ 95% of samples yielded positive results. (Refer to Table 7 for values).
Inclusivity Study:
- Study Type: Analytical reactivity evaluation.
- Sample Size: Panels of eight seasonal influenza A H1N1 strains, 10 seasonal influenza A H3N2 strains, and 11 2009 H1N1 influenza strains.
- Key Results: Most strains detected at or near LoD. Four of 29 influenza strains not detected. Variability in Flu A H1 assay detection for some strains. Specific H3N2 strains not detected (A/Aichi/2/68, A/Hong Kong/8/68, A/MRC-2 recomb).
Exclusivity Study:
- Study Type: Cross-reactivity potential evaluation.
- Sample Size: Simulated NPS samples with high concentrations of non-target influenza viruses, bacteria, and non-influenza viruses.
- Key Results: Assays gave expected negative results with non-target influenza assays and non-influenza exclusivity panel organisms. Exceptions: Flu A Sw assay detected three H1N1 swine-lineage strains; Flu A H3 and Flu A Sw assays detected A/SW/IA/1/99 (swine origin H3N2).
Reproducibility Study:
- Study Type: Multicenter study.
- Sample Size: Six total panels of NPS and NPW samples for each of three influenza types (A H1N1, A H3N2, 2009 H1N1 influenza). Each panel had 9 samples (high negative, low positive, medium positive). Tested twice daily at three sites (30 results per sample, 90 results per spike level).
- Key Results: Detection rate was ≥ 98% for samples containing influenza virus ≥ LoD. Samples spiked below LoD had variable results. (Refer to Table 13, Table 14, and Table 15 for detailed results for Flu A H1, Flu A H3, and combined Flu A H1 2009 and Flu A Sw assays).
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
- Positive Percent Agreement (PPA)
- Negative Percent Agreement (NPA)
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.
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.
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
§ 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.
0
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.
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
2
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.
3
| 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 A | ||
| Subtyping Kit | CDC rRT-PCR Flu Panel | ||
| (K080570) | CDC rRT-PCR 2009 | ||
| A(H1N1)pdm Flu Panel | |||
| (K101564) | |||
| Viruses Detected | Influenza A/H1, Influenza A/H3 | ||
| and Influenza A/2009 H1N1 | Influenza A, Influenza B, | ||
| Influenza A/H1, Influenza A/H3 | |||
| and Influenza A/H5 | Influenza A and Influenza A | ||
| 2009 H1N1 | |||
| Specimen types | Nasopharyngeal swabs, | ||
| nasopharyngeal washes | Nasopharyngeal swabs, nasal | ||
| swabs, and virus culture | Nasopharyngeal swabs, nasal | ||
| swabs, nasal aspirates, nasal | |||
| washes, dual nasopharyngeal / | |||
| throat swabs, broncheoalveolar | |||
| lavage, tracheal aspirate, | |||
| bronchial wash and viral culture | |||
| Required | |||
| Instrumentation | JBAIDS instrument | Applied Biosystems 7500 Fast | |
| Dx Real-time PCR instrument | |||
| with SDS software v 1.4 | Applied Biosystems 7500 Fast | ||
| Dx Real-time PCR instrument | |||
| Interpretation of | |||
| Test Results | Automated analysis of test | ||
| results and controls | User required to interpret test | ||
| and control results | User required to interpret test | ||
| and control results | |||
| Enzyme Master | |||
| Mix | Assays come in freeze-dried | ||
| single use vials that include all | |||
| components of master mix | Invitrogen SuperScript™ III | ||
| Platinum® One-Step | |||
| Quantitative RT-PCR Kits | Invitrogen SuperScript™ III | ||
| Platinum® One-Step | |||
| Quantitative RT-PCR Kits | |||
| Reagent Storage | Reagents are stored at room | ||
| temperature | Reagents are stored at ≤ -20°C | Reagents are stored at ≤ -20°C | |
| Extraction | |||
| Methods | • IT 1 - 2 -3™ Platinum Path | ||
| Sample Purification Kit | • Qiagen QIAamp® Viral RNA | ||
| Mini Kit | • Qiagen QIAamp® Viral RNA | ||
| Mini Kit | |||
| • Roche MagNA Pure Compact | |||
| Nucleic Acid Isolation Kit I | • Qiagen RNeasy® Mini Kit | ||
| • Roche MagNA Pure TNA Kit | • Roche MagNA Pure Compact | ||
| Nucleic Acid Isolation Kit I | |||
| Roche MagNA Pure TNA Kit | |||
| • Roche MagNA Pure LC RNA | |||
| Isolation Kit II | • Roche MagNA Pure LC RNA | ||
| isolation Kit II | |||
| • Qiagen QIAcube with | |||
| QIAamp viral RNA mini kit | |||
| • bioMerieux NucliSENS | |||
| easyMAG |
:
.
Table 1. Similarities Between the JBADS Influenza A Subtyping Kit and its Predicate Devices
x
.
,
·
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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,
5
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 | 206 | ||||
| ર્સ | 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%) | |
| Age | |||||||
| Range® | 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
6
two-sided 95% confidence interval was calculated. The results are summarized in Table 4.
| Influenza A
Strain | Sample
Matrix | Purification
Kit | 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
7
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.
| Influenza
Assay | Sample
Type | 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
8
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 2009 | |||
| and 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.
9
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 | Lowest
Concentration
Detected |
|--------------------------|-------------------------------------|
| 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 Concentration
Detected |
|----------------------|----------------------------------|
| 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
10
| Strain | Lowest Concentration
Detected |
|--------------------------|----------------------------------|
| 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 | Concentration
Tested | Assays
Tested |
|-------------|-----------------------------------|--------------------------------------|-------------------------|------------------|
| 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-Human
Recombinant) | A/Vietnam/1203/2004(H5N1)-PR8 | 3.16E+07 EID50/mL | Flu A H1
2009 |
| | 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 |
11
| Type/Subtype | Strain | Concentration
Tested | Assays
Tested |
|---------------------------------------------|----------------------------|--------------------------------------------------|------------------------------|
| 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 of
swine lineage) | A/Maryland/12/1991 | 1.00E+05 TCID50/mL | |
| H1N1 (Human of
swine lineage) | A/Iowa/1/2006 | 1.00E+05 TCID50/mL | |
| H7N2 (Human) | A/New York/107/2003 | 30 µl of an unknown
concentration into
1mL | |
| | 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 H1
2009 |
| | 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 A
H1N1 | A/NWS/33 | 4.24E+03 TCID50/mL | Flu A Sw |
| | A1/Denver/1/57 | 4.24E+03 TCID50/mL | Flu A H1
2009 |
| | 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 A
H3N2 | A/Hong Kong/8/68 | 1.00E+05 TCID50/mL | Flu A Sw
Flu A H1
2009 |
| | 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 | Concentration
Tested | Assays
Tested |
| | 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 | |
12
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,
13
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.
| 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
(% Pos.) | 95% CI | |
|----------------|-------------------------|---------------------------------------------------|-----------------|-----------------|----------------------------------------------------------|-----------------|-----------------|-----------------|-------------------------------------|------------------|-----------|
| | | Site 1 | Site 2 | Site 3 | Overall
for All
Sites | Site 1 | Site 2 | Site 3 | Overall
for All
Sites | | |
| 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 |
| | Detection
all 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 |
| | Detection
all 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)
14
| | | IT 1-2-3 Platinum Path
Sample Purification Kit | | | | Roche MagNA Pure Compact
Nucleic Acid Isolation Kit I | | | | | |
|----------------|-------------------------|---------------------------------------------------------------------|-----------------|-----------------|-----------------------------|---------------------------------------------------------------------|-----------------|-----------------|-----------------------------|-------------------------------------|-----------|
| Sample
Type | Virus
Spike
Level | 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 | 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%) | 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 |
| | Detection
all 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 |
| | Detection
all 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.
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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:
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.
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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.
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