(105 days)
The Alere™ i Influenza A & B 2 assay performed on the Alere™ i Instrument is a rapid molecular in vitro diagnostic test utilizing an isothermal nucleic acid amplification technology for the qualitative detection and discrimination of influenza A and B viral RNA in direct nasal or nasopharyngeal swabs and nasal or nasopharyngeal swabs eluted in viral transport media from patients with signs and symptoms of respiratory infection. It is intended for use as an aid in the differential diagnosis of influenza A and B viral infections in humans in conjunction with clinical and epidemiological risk factors. The assay is not intended to detect the presence of influenza C virus.
Negative results do not preclude influenza virus infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.
Performance characteristics for influenza A were established during the 2016-2017 influenza season when influenza A/H3 and A/H1N1 pandemic were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characterics may vary.
If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to state or local health department for testing. Viral culture should not be attempted in cases unless a BSL 3+ facility is available to receive and culture specimens.
Alere™ i Influenza A & B 2 is a rapid, instrument-based isothermal test for the qualitative detection and differentiation of influenza A and influenza B from nasal swabs tested directly or after elution in viral transport media collected from patients with signs and symptoms of respiratory infection. The Alere™ i Influenza A & B 2 System utilizes isothermal nucleic acid amplification technology and is comprised of:
- Sample Receiver single use, disposable containing the elution buffer
- Test Base single use, disposable comprising two sealed reaction tubes, each containing a lyophilized pellet
- Transfer Cartridge – Single use, disposable for transfer of the eluted sample to the Test Base, and
- Alere™ i Instrument - repeat use reader
The reaction tubes in the Test Base contain the reagents required for amplification of the target nucleic acid and an internal control. Alere™ i Influenza A & B 2 utilizes a pair of templates (similar to primers) for the specific amplification of RNA from influenza A and B, which occur in two separate reaction tube contains fluorescently labeled molecular beacon designed to specifically identify the amplified RNA targets. Alere™ i Influenza A & B 2 is performed within the confinement of the Test Base, and no other part of the Alere™ i Instrument has contact with the sample during the amplification process. This minimizes the risk of instrument contamination and sample carry-over between measurements.
To perform the assay, the Sample Receiver and Test Base are inserted into the Alere™ i Instrument and the elution buffer is automatically heated by the instrument. The sample is added to the Sample Receiver and transferred via the Transfer Cartridge to the Test Base, re-suspending the lyophilized pellets contained within the Test Base and initiating target amplification. Heating, mixing and detection by fluorescence are provided by the instrument, with results automatically reported.
Results are displayed by the Alere™ i Instrument and are also stored in an on-board archive and are assigned to a sample ID that has been entered into the Alere™ i Instrument by the operator, and the date/time the test was performed. Data can be retrieved and downloaded by the operator at any time after testing. An external Alere™ Universal Printer can be attached via USB to the Alere™ i Instrument to print test results.
Here's an analysis of the acceptance criteria and study detailed in the provided text:
1. Table of Acceptance Criteria and Reported Device Performance
The document does not explicitly state pre-defined acceptance criteria (e.g., "the device must achieve X sensitivity and Y specificity"). Instead, it presents the device's performance, which is implicitly accepted by the FDA's "Substantially Equivalent" determination. The table below summarizes the reported clinical performance.
| Criterion (Implicit Acceptance) | Reported Device Performance (Direct Nasal/Nasopharyngeal Swab) | Reported Device Performance (Nasal/Nasopharyngeal Swab Eluted in VTM) |
|---|---|---|
| Influenza A Sensitivity | 96.3% (95%CI: 93.3%-98.2%) | 92.8% (95%CI: 89.0%-95.6%) |
| Influenza A Specificity | 97.4% (95%Cl: 96.0%-98.4%) | 98.5% (95%CI: 97.4%-99.2%) |
| Influenza B Sensitivity | 100% (95%CI: 96.3%-100%) | 100% (95%CI: 96.3%-100%) |
| Influenza B Specificity | 97.1% (95%CI: 95.9%-98.1%) | 97.7% (95%CI: 96.6%-98.6%) |
| Direct Swab Invalid Rate (after re-testing) | 0.4% (95% Cl: 0.1% to 1.0%) | N/A |
| VTM Invalid Rate (after re-testing) | N/A | 1.0% (95% Cl: 0.6%) |
| Analytical Sensitivity (LoD) (e.g., A/Texas/50/2012 A/H3N2 Direct Swab) | 1.00 x 10^-1 TCID50/mL | 1.00 x 10^0 TCID50/mL |
| Analytical Sensitivity (LoD) (e.g., A/Texas/50/2012 A/H3N2 VTM) | N/A | 1.00 x 10^0 TCID50/mL |
| Analytical Reactivity (Inclusivity) | Positive detection across various influenza A and B strains at specified concentrations (see document for full list) | Positive detection across various influenza A and B strains at specified concentrations (see document for full list) |
| Analytical Specificity (Cross-Reactivity) | No cross-reactivity observed for 36 tested microorganisms (with minor exceptions for E. coli, Moraxella catarrhalis, and Proteus vulgaris at high concentrations) | No cross-reactivity observed for 36 tested microorganisms (with minor exceptions for E. coli, Moraxella catarrhalis, and Proteus vulgaris at high concentrations) |
| Interfering Substances | No adverse effect on test performance with listed substances | No adverse effect on test performance with listed substances |
| Reproducibility | 100% agreement for moderate positive Influenza A and B; 98.9% for low positive Influenza B. All true negatives were negative. | 100% agreement for moderate positive Influenza A and B; 98.9% for low positive Influenza B. All true negatives were negative. |
2. Sample Size Used for the Test Set and Data Provenance
- Sample Size:
- Total enrolled: 1110 nasal or nasopharyngeal swab specimens.
- Total tested:
- 1070 for direct swab performance analysis (after excluding 36 not meeting eligibility and 4 invalid after re-testing).
- 1057 for viral transport media (VTM) performance analysis (after excluding 36 not meeting eligibility, 11 invalid after re-testing, and 6 not meeting eligibility).
- Data Provenance:
- Country of Origin: United States (multi-center, conducted at ten US trial sites).
- Retrospective or Prospective: Prospective clinical study.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts
The ground truth for the clinical study was established using an FDA-cleared real-time Polymerase Chain Reaction (RT-PCR) test as the comparator method. The document does not mention the use of human experts (e.g., radiologists) to establish ground truth for this device, as it is an in vitro diagnostic test for viral RNA detection, not an imaging device.
4. Adjudication Method for the Test Set
The ground truth was established by a single comparator method (FDA-cleared RT-PCR). There is no mention of an adjudication method for the test set, as the RT-PCR result was considered the definitive truth. For discrepancies, secondary molecular testing was used to investigate false positives/negatives (e.g., "Flu A nucleic acid was detected in 6/21 False positive specimens using a second FDA-cleared molecular test"). This refers to investigation of the Alere™ i results against the initial RT-PCR, not an adjudication process to establish the initial ground truth.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
No MRMC comparative effectiveness study was done. This type of study is typically relevant for interpretative devices (e.g., medical imaging AI) where human readers are involved in the diagnostic process. The Alere™ i Influenza A & B 2 is an automated molecular diagnostic test with automated result interpretation.
6. Standalone Performance
Yes, a standalone performance study was done. The clinical study results (sensitivity, specificity, invalid rates) reported for the device directly reflect its performance as an algorithm-only (kit + instrument) system without human-in-the-loop interpretation beyond operating the instrument and reading the automated result. The "automated result interpretation" listed in the device comparison table further confirms its standalone nature.
7. Type of Ground Truth Used
The primary ground truth used for the clinical performance evaluation was an FDA-cleared real-time Polymerase Chain Reaction (RT-PCR) test. This is a molecular diagnostic method considered highly accurate for detecting viral RNA. Secondary molecular testing was used to investigate discordant results.
8. Sample Size for the Training Set
The document does not specify the sample size for the training set. It details the clinical validation (test set) and analytical studies. Typically, for such devices, the training data would be proprietary to the manufacturer and not explicitly disclosed in an FDA 510(k) summary, which focuses on validation data.
9. How the Ground Truth for the Training Set Was Established
Since the training set data is not disclosed, the method for establishing its ground truth is also not described in this document. It is generally assumed that the training data for molecular diagnostic assays would be similarly characterized using highly accurate reference methods, potentially including PCR, sequencing, or well-characterized viral cultures.
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Image /page/0/Picture/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo is a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is a stylized image of three human profiles facing to the right, with flowing lines representing hair or movement.
September 29, 2017
Public Health Service
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
Alere Scarborough, Inc. Angela Drysdale VP, Regulatory Affairs - Infectious Disease 10 Southgate Road Scarborough ME 04074
Re: K171792
Trade/Device Name: Alere i Influenza A & B 2. Alere i Instrument, Alere i Influenza A & B Control Swab Kit Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: II Product Code: OCC, OZE, OOI Dated: June 15, 2017 Received: July 3, 2017
Dear Ms. Drysdale:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) 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 regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours,
Steven R. Gitterman -S
for
Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known) K171792
Device Name Alere i Influenza A & B 2
Indications for Use (Describe)
The Alere i Influenza A & B 2 assay performed on the Alere i Instrument is a rapid molecular in vitro diagnostic test utilizing an isothermal nucleic acid amplification technology for the qualitative detection and discrimination of influenza A and B viral RNA in direct nasal or nasopharyngeal swabs and nasal or nasopharyngeal swabs eluted in viral transport media from patients with signs and symptoms of respiratory infection. It is intended for use as an aid in the differential diagnosis of influenza A and B viral infections in humans in conjunction with clinical and epidemiological risk factors. The assay is not intended to detect the presence of influenza C virus.
Negative results do not preclude influenza virus infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.
Performance characteristics for influenza A were established during the 2016-2017 influenza season when influenza A/H3 and A/H1N1 pandemic were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characterics may vary.
If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to state or local health department for testing. Viral culture should not be attempted in cases unless a BSL 3+ facility is available to receive and culture specimens.
Type of Use (Select one or both, as applicable)
| Prescription Use (Part 21 CFR 801 Subpart D) | |
|---|---|
| Over-The-Counter Use (21 CFR 801 Subpart C) |
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510(k) SUMMARY
This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.
The assigned 510(k) number is: K171792
SUBMITTER
Alere Scarborough, Inc. 10 Southgate Road Scarborough, Maine 04074 Establishment Registration Number: 1221359
CONTACT PERSON
Angela Drysdale (207) 415-1393 (office) (207) 730-5767 (FAX) angela.drysdale@alere.com (email)
DATE PREPARED
June 15, 2017
TRADE NAME Alere™ i Influenza A & B 2
COMMON NAME
Alere™ i Flu, Alere™ i, Alere™ i Flu 2, Alere™ Influenza A & B, Alere™ Influenza A & B 2
CLASSIFICATION NAME
Respiratory Viral Panel Multiplex Nucleic Acid System (per 21 CFR 866.3980) Instrumentation for Clinical Multiplex Test Systems (per 21 CFR 862.2570)
CLASSIFICATION Class II
PRODUCT CODE OCC, OZE, OOI
PANEL Microbiology (83)
PREDICATE DEVICE Alere™ i Influenza A & B K163266
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DEVICE DESCRIPTION
Alere™ i Influenza A & B 2 is a rapid, instrument-based isothermal test for the qualitative detection and differentiation of influenza A and influenza B from nasal swabs tested directly or after elution in viral transport media collected from patients with signs and symptoms of respiratory infection. The Alere™ i Influenza A & B 2 System utilizes isothermal nucleic acid amplification technology and is comprised of:
- Sample Receiver single use, disposable containing the elution buffer ●
- Test Base single use, disposable comprising two sealed reaction tubes, each containing a lyophilized ● pellet
- . Transfer Cartridge – Single use, disposable for transfer of the eluted sample to the Test Base, and
- Alere™ i Instrument - repeat use reader
The reaction tubes in the Test Base contain the reagents required for amplification of the target nucleic acid and an internal control. Alere™ i Influenza A & B 2 utilizes a pair of templates (similar to primers) for the specific amplification of RNA from influenza A and B, which occur in two separate reaction tube contains fluorescently labeled molecular beacon designed to specifically identify the amplified RNA targets. Alere™ i Influenza A & B 2 is performed within the confinement of the Test Base, and no other part of the Alere™ i Instrument has contact with the sample during the amplification process. This minimizes the risk of instrument contamination and sample carry-over between measurements.
To perform the assay, the Sample Receiver and Test Base are inserted into the Alere™ i Instrument and the elution buffer is automatically heated by the instrument. The sample is added to the Sample Receiver and transferred via the Transfer Cartridge to the Test Base, re-suspending the lyophilized pellets contained within the Test Base and initiating target amplification. Heating, mixing and detection by fluorescence are provided by the instrument, with results automatically reported.
Results are displayed by the Alere™ i Instrument and are also stored in an on-board archive and are assigned to a sample ID that has been entered into the Alere™ i Instrument by the operator, and the date/time the test was performed. Data can be retrieved and downloaded by the operator at any time after testing. An external Alere™ Universal Printer can be attached via USB to the Alere™ i Instrument to print test results.
INTENDED USE
The Alere™ i Influenza A & B 2 assay performed on the Alere™ i Instrument is a rapid molecular in vitro diagnostic test utilizing an isothermal nucleic acid amplification technology for the qualitative detection and discrimination of influenza A and B viral RNA in direct nasal or nasopharyngeal swabs and nasal or nasopharyngeal swabs eluted in viral transport media from patients with signs and symptoms of respiratory infection. It is intended for use as an aid in the differential diagnosis of influenza A and B viral infections in conjunction with clinical and epidemiological risk factors. The assay is not intended to detect the presence of influenza C virus.
Negative results do not preclude influenza virus infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.
Performance characteristics for influenza A were established during the 2016-2017 influenza season when influenza A/H3 and A/H1N1 pandemic were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary.
If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to state or local health department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
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TECHNICAL CHARACTERISTICS
Alere™ i Influenza A & B 2 and the predicate device, Alere™ i Influenza A & B, have the same intended use, indications for use, and utilize similar basic principles of operation. They are both molecular tests for the qualitative detection of influenza A and influenza B viral RNA.
DEVICE COMPARISON
Alere™ i Influenza A & B 2 was compared to the legally marketed predicate device, the Alere™ i Influenza A & B Assay.
| Parameter | Alere™ i Influenza A & B 2 | Alere™ i Influenza A & B(K163266) |
|---|---|---|
| FDA Product Code | OCC, OZE, OOI | Same |
| Assay Target | Influenza A, Influenza B | Same |
| Intended Use | The Alere™ i Influenza A & B 2 assay performedon the Alere™ i Instrument is a rapid molecular in vitro diagnostic test utilizing an isothermal nucleicacid amplification technology for the qualitativedetection and discrimination of influenza A and Bviral RNA in direct nasal or nasopharyngealswabs and nasal or nasopharyngeal swabseluted in viral transport media from patients withsigns and symptoms of respiratory infection. It isintended for use as an aid in the differentialdiagnosis of influenza A and B viral infections inhumans in conjunction with clinical andepidemiological risk factors. The assay is notintended to detect the presence of influenza Cvirus. | The Alere™ i Influenza A & B assay performedon the Alere™ i Instrument is a rapid molecular in vitro diagnostic test utilizing an isothermalnucleic acid amplification technology for thequalitative detection and discrimination ofinfluenza A and B viral RNA in direct nasalswabs and nasal or nasopharyngeal swabseluted in viral transport media from patientswith signs and symptoms of respiratoryinfection. It is intended for use as an aid in thedifferential diagnosis of influenza A and B viralinfections in humans in conjunction with clinicaland epidemiological risk factors. The assay isnot intended to detect the presence ofinfluenza C virus. |
| Negative results do not preclude influenza virusinfection and should not be used as the solebasis for diagnosis, treatment or other patientmanagement decisions. | Negative results do not preclude influenza virusinfection and should not be used as the solebasis for diagnosis, treatment or other patientmanagement decisions. | |
| Performance characteristics for influenza A wereestablished during the 2016-2017 influenzaseason when influenza A/H3 and A/H1N1pandemic were the predominant influenza Aviruses in circulation. When other influenza Aviruses are emerging, performancecharacteristics may vary. | Performance characteristics for influenza Awere established during the 2012-2013 and the2014-2015 influenza seasons when influenzaA/H3 and A/H1N1 pandemic were thepredominant influenza A viruses in circulation.When other influenza A viruses are emerging,performance characteristics may vary. | |
| If infection with a novel influenza A virus issuspected based on current clinical andepidemiological screening criteria recommendedby public health authorities, specimens should becollected with appropriate infection controlprecautions for novel virulent Influenza virusesand sent to state or local health department fortesting. Viral culture should not be attempted inthese cases unless a BSL 3+ facility is availableto receive and culture specimens. | If infection with a novel influenza A virus issuspected based on current clinical andepidemiological screening criteriarecommended by public health authorities,specimens should be collected with appropriateinfection control precautions for novel virulentInfluenza viruses and sent to state or localhealth department for testing. Viral cultureshould not be attempted in these cases unlessa BSL 3+ facility is available to receive andculture specimens. | |
| 2Intended Environmentfor Use | Professional use, in a medical laboratory or point-of-care | Same |
| Instrumentation | Alere™ i Instrument | Same |
| Self-Contained System | Integrated PC, Software and Touch ScreenDisplay | Same |
| Automated Assay | Yes. Sample preparation, amplification, detection, and result interpretation. | Same |
| Assay Information | ||
| Sample Type | Nasopharyngeal Swab, Nasopharyngeal Swab | Nasal Swab, Nasal Swab or Nasopharyngeal |
| Parameter | Alere™ i Influenza A & B 2 | Alere™ i Influenza A & B(K163266) |
| eluted in Viral Transport Media, Nasal Swab,Nasal Swab eluted in Viral Transport Media | Swab eluted in Viral Transport Media | |
| Influenza A Target | PB2 Segment | Same |
| Influenza B Target | PA Segment | Same |
| Technology | Isothermal nucleic acid amplification for detectingthe presence/absence of viral RNA in clinicalspecimens. | Same |
| Internal Control | Yes | Same |
| Results Interpretation | Automated | Same |
| Assay Result | Qualitative | Same |
| Time to Result | <15 minutes | Same |
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PERFORMANCE SUMMARY
CLINICAL STUDY
The clinical performance of Alere™ i Influenza A & B 2 was established in a multi-center, prospective clinical study conducted at ten US trial sites during the 2016-2017 respiratory season.
A total of 1110 nasal or nasopharyngeal swab specimens were enrolled in this study. Of those, 36 nasal or nasopharyngeal swab specimens did not meet eligibility criteria. A total of 1074 specimens were tested with Alere™ i Influenza A & B 2. 56% of the population tested was female and 44% was male.
In this study, two nasopharyngeal swabs were collected from each Subject. One swab was tested directly with Alere™ i Influenza A & B 2, according to product instructions for testing direct swabs. The other swab was eluted in VTM, and a sample of the VTM eluate was tested with Alere™ i Influenza A & B 2. An FDA-cleared real-time Polymerase Chain Reaction (RT-PCR) test was utilized as the comparator method for this study.
Of the 1074 specimens. Alere™ i Influenza A & B 2 generated invalid results for 4 direct swab specimens after repeat testing per the product instructions, resulting in a total of 1070 specimens for direct swab performance analysis. Alere™ i Influenza A & B 2 generated invalid results for 11 viral transport media specimens after repeat testing per the product instructions and an additional 6 specimens did not meet eligibility criteria, resulting in a total of 1057 specimens for viral transport media performance analysis.
Compared to the comparator method, the performance of Alere™ i Influenza A & B 2 is presented in the tables below.
Direct Nasal or Nasopharyngeal Swab – Performance Obtained for Influenza A with Alere™ i Influenza A & B 2 against the Comparator Method
| Alere™ i | Comparator Method | ||
|---|---|---|---|
| Influenza A & B 2- Flu A | Positive | Negative | Total |
| Positive | 260 | 21a | 281 |
| Negative | 10b | 779 | 789 |
| Total | 270 | 800 | 1070 |
| Sensitivity: 260/270 | 96.3% | (95%CI: 93.3%-98.2%) | |
| Specificity: 779/800 | 97.4% | (95%Cl: 96.0%-98.4%) |
ª Flu A nucleic acid was detected in 6/21False positive specimens using a second FDA-cleared molecular test
ʰ Flu A nucleic acid was not detected in 4/10 False negative specimens using a second FDA-cleared molecular test
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Direct Nasal or Nasopharyngeal Swab – Performance Obtained for Influenza B with Alere™ i Influenza A & B 2 against the Comparator Method
| Alere™ iInfluenza A & B 2- Flu B | Comparator Method | ||
|---|---|---|---|
| Positive | Negative | Total | |
| Positive | 97 | 28a | 125 |
| Negative | 0 | 945 | 945 |
| Total | 97 | 973 | 1070 |
| Sensitivity: | 97/97100% | (95%CI: 96.3%-100%) | |
| Specificity: | 945/97397.1% (95%CI: 95.9%-98.1%) |
ª Flu B nucleic acid was detected in 21/28 False positive specimens using a second FDA-cleared molecular test
Nasal or Nasopharyngeal Swab Eluted in Viral Transport Media – Performance Obtained for Influenza A with Alere™ i Influenza A & B 2 against the Comparator Method
| Alere™ iInfluenza A & B 2- Flu A | Comparator Method | ||
|---|---|---|---|
| Positive | Negative | Total | |
| Positive | 246 | 12a | 258 |
| Negative | 19b | 780 | 799 |
| Total | 265 | 792 | 1057 |
| Sensitivity: 246/265 | 92.8% | (95%CI: 89.0%-95.6%) | |
| Specificity: 780/792 | 98.5% | (95%CI: 97.4%-99.2%) |
a Flu A nucleic acid was detected in 5/12 False positive specimens using a second FDA-cleared molecular test
° Flu A nucleic acid was not detected 6/19 in False negative specimens using a second FDA-cleared molecular test
Nasal or Nasopharyngeal Swab Eluted in Viral Transport Media – Performance Obtained for Influenza B with Alere™ i Influenza A & B 2 against the Comparator Method
| Alere™ i | Comparator Method | ||
|---|---|---|---|
| Influenza A & B 2- Flu B | Positive | Negative | Total |
| Positive | 97 | 22a | 119 |
| Negative | 0 | 938 | 938 |
| Total | 97 | 960 | 1057 |
| Sensitivity: 97/97 | 100% | (95%CI: 96.3%-100%) | |
| Specificity: 938/960 | 97.7% | (95%CI: 96.6%- 98.6%) |
ª Flu B nucleic acid was detected in 18/22 False positive specimens using a second FDA-cleared molecular test
During the prospective clinical study, the initial invalid rate for direct nasal or nasopharyngeal swab samples (before repeat testing per the product instructions) was 0.8% (9/1074) (95% Cl: 0.4% to 1.6%). After repeat testing per the product instructions, the invalid rate was 0.4% (4/1074) (95% Cl: 0.1% to 1.0%).
The initial invalid rate for swabs eluted in viral transport media was 2.2% (24/1074) (95% Cl: 1.5% to 3.2%). After repeat testing per the product instructions, the invalid rate was 1.0% (11/1074) (95% Cl: 0.6%).
ANALYTICAL STUDIES
ANALYTICAL SENSITIVITY
Alere™ i Influenza A & B 2 limit of detection (LOD or Cos), defined as the concentration of influenza A or B that produces positive Alere™ i Influenza A & B 2 results approximately 95% of the time, was identified by evaluating two influenza A strains and two influenza B strains for both direct swab and swab eluted in VTM testing in Alere™ i Influenza A & B 2. The concentrations identified as the LOD (or Coss) levels for each strain and testing method are listed below.
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| Influenza Strain | Influenza ASubtype orInfluenza B GeneticLineage | LoD(TCID50/mL) | LoD(TCID50/Swab)* | LoD (GenomeEquivalents/mL) | LoD (GenomeEquivalents/Swab)* |
|---|---|---|---|---|---|
| A/Texas/50/2012 | A/H3N2 | $1.00 x 10^{-1}$ | $1.00 x 10^{-3}$ | $1.06 x 10^{4}$ | $1.06 x 10^{2}$ |
| A/California/7/2009 | A/2009 H1N1 pdm | $2.00 x 10^{0}$ | $2.00 x 10^{-2}$ | $1.60 x 10^{4}$ | $1.60 x 10^{2}$ |
| B/Brisbane/60/2008 | Victoria lineage | $5.20 x 10^{1}$ | $5.20 x 10^{-1}$ | $6.60 x 10^{3}$ | $6.60 x 10^{1}$ |
| B/Wisconsin/1/2010 | B Yamagata lineage | $5.01 x 10^{2}$ | $5.01 x 10^{0}$ | $1.11 x 10^{4}$ | $1.11 x 10^{2}$ |
Limit of Detection (LOD) Study Results – Natural Nasal Swab Matrix (Direct Swab Testing)
*Note: 10 µl of each virus dilution was coated onto a swab
Limit of Detection (LOD) Study Results – Natural Nasal Swab Matrix (Swab Eluted in VTM Testing)
| Influenza Strain | Influenza ASubtype orInfluenza BGenetic Lineage | LoD(TCID50/mL) | LOD(TCID50/Swab)* | LoD (GenomeEquivalents/mL | LoD (GenomeEquivalents/Swab)* |
|---|---|---|---|---|---|
| A/Texas/50/2012 | A/H3N2 | 1.00 x 10° | 1.00 x 10-2 | 2.10 x 10° | 2.10 x 10° |
| A/California/7/2009 | A/2009 H1N1 pdm | 5.00 x 101 | 5.00 x 10.7 | 3.83 x 10° | 3.83 x 103 |
| B/Brisbane/60/2008 | Victoria lineage | 1.20 x 10° | 1.20 x 10 | 1.51 x 10° | 1.51 x 10° |
| B/Wisconsin/1/2010 | B Yamagatalineage | 9.66 x 103 | 9.66 x 10 | 2.14 x 10° | 2.14 x 10° |
*Note: 10 µl of each virus dilution was coated onto a swab; each contrived swab was further diluted into 3 mL of UTM
ANALYTICAL REACTIVITY (INCLUSIVITY)
The influenza A and B strains listed below were tested and generated positive Alere™ i Influenza A & B 2 test results at the concentrations indicated in the table.
| Influenza ASubtype orInfluenza BGenetic Lineage | Test Concentration (in TCID50 or Genome Equivalents) | ||||
|---|---|---|---|---|---|
| Influenza Strain | TCID50/mL | TCID50/Swab* | GenomeEquivalents/mL | GenomeEquivalents/Swab* | |
| A/New Caledonia/20/1999 | A/H1N1 | $2.74 x 10^4$ | $2.74 x 10^2$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/New Jersey/8/76 | A/H1N1 | $8.62 x 10^{-1}$ | $8.62 x 10^{-3}$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Brisbane/59/2007 | A/H1N1 | $2.44 x 10^0$ | $2.44 x 10^{-2}$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/WSN/33 | A/H1N1 | $2.78 x 10^2$ | $2.78 x 10^0$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/California/4/2009 | A/H1N1 | $1.26 x 10^1$ | $1.26 x 10^1$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Maryland/04/2011 | A/H1N1 | $1.55 x 10^3$ | $1.55 x 10^1$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/New York/18/2009 | A/H1N1 | $9.08 x 10^0$ | $9.08 x 10^{-2}$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/South Carolina/2/2010 | A/H1N1 | $3.47 x 10^1$ | $3.47 x 10^{-1}$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Port Chalmers/1/73 | A/H3N2 | $2.02 x 10^3$ | $2.02 x 10^1$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Hong Kong/8/68 | A/H3N2 | $2.16 x 10^1$ | $2.16 x 10^{-1}$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Aichi/2/68 | A/H3N2 | $3.58 x 10^0$ | $3.58 x 10^{-2}$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Perth/16/2009 | A/H3N2 | $6.12 x 10^2$ | $6.12 x 10^0$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Victoria/3/75 | A/H3N2 | $9.61 x 10^{-1}$ | $9.61 x 10^{-3}$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Wisconsin/67/2005 | A/H3N2 | $1.60 x 10^3$ | $1.60 x 10^1$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Brisbane/10/2007 | A/H3N2 | $5.48 x 10^1$ | $5.48 x 10^{-1}$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Victoria/361/2011 | A/H3N2 | $6.41 x 10^0$ | $6.41 x 10^{-2}$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Indiana/10/2011 | A/H3N2v | $7.02 x 10^3$ | $7.02 x 10^1$ | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Sichuan/26221/2014 (inactivated) | A/H5N6 | N/A | N/A | $3.00 x 10^4$ | $3.00 x 10^2$ |
| A/Anhui/1/2013 (inactivated) | A/H7N9 | N/A | N/A | $6.70 x 10^4$ | $6.70 x 10^2$ |
| B/Lee/40 | Victoria Lineage | $1.60 x 10^0$ | $1.60 x 10^{-2}$ | $2.25 x 10^4$ | $2.25 x 10^2$ |
| B/Victoria/504/2000 | Victoria Lineage | $1.45 x 10^2$ | $1.45 x 10^0$ | $2.25 x 10^4$ | $2.25 x 10^2$ |
| B/Nevada/03/2011 | Victoria Lineage | $3.66 x 10^2$ | $3.66 x 10^0$ | $2.25 x 10^4$ | $2.25 x 10^2$ |
| B/Montana/05/2012 | Victoria Lineage | $2.21 x 10^2$ | $2.21 x 10^0$ | $2.25 x 10^4$ | $2.25 x 10^2$ |
| B/Maryland/1/59 | Yamagata Lineage | $8.42 x 10^2$ | $8.42 x 10^0$ | $2.25 x 10^4$ | $2.25 x 10^2$ |
| B/Russia/69 | Yamagata Lineage | $9.38 x 10^2$ | $9.38 x 10^0$ | $7.23 x 10^5$ | $7.23 x 10^3$ |
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| B/Bangladesh/3333/2007 | Yamagata Lineage | $4.64 x 10^2$ | $4.64 x 10^0$ | $2.33 x 10^4$ | $2.33 x 10^2$ |
|---|---|---|---|---|---|
| B/Massachusetts/2/2012 | Yamagata Lineage | $4.30 x 10^2$ | $4.30 x 10^0$ | $2.25 x 10^4$ | $2.25 x 10^2$ |
| B/Malaysia/2506/2004 | Yamagata Lineage | $3.25 x 10^3$ | $3.25 x 10^1$ | $2.25 x 10^4$ | $2.25 x 10^2$ |
| B/Texas/06/2011 | Yamagata Lineage | $5.33 x 10^2$ | $5.33 x 10^0$ | $2.25 x 10^4$ | $2.25 x 10^2$ |
*Note: 10 µl of each virus dilution was coated onto a swab
ANALYTICAL SPECIFICITY (CROSS-REACTIVITY)
To determine the analytical specificity of Alere™ i Influenza A & B 2, 36 commensal and pathogenic microorganisms (18 bacteria, 17 viruses and 1 yeast) that may be present in the nasal cavity or nasopharynx were tested. All of the following microorganisms were negative when tested at concentrations ranging from 10° to 10° cells/mL or CFU/mL (bacteria), 10 to 10° TCID50mL (viruses), and 10° cells/mL (yeast).
| Bacteria | Viruses | Yeast |
|---|---|---|
| Bordetella pertussis | Adenovirus type 1 | Candida albicans |
| Corynebacterium diphtheriae | Adenovirus type 7 | |
| Escherichia coli* | Human Coronavirus OC43 | |
| Haemophilius influenzae | Echovirus 7 | |
| Klebsiella pneumoniae | Human Coronavirus 229E | |
| Lactobacillus plantarum | Enterovirus 70 | |
| Legionella pneumophila | Coxsackievirus B4 | |
| Moraxella/Branhamella catarrhalis* | Human Cytomegalovirus (CMV) (HerpesV) | |
| Mycobacterium tuberculosis | Human metapneumovirus | |
| Mycoplasma pneumoniae | Rhinovirus 1A | |
| Neisseria meningitidis | Measles (Edmonston) | |
| Proteus vulgaris* | Mumps (Enders) | |
| Pseudomonas aeruginosa | Parainfluenza 1 | |
| Staphylococcus aureus | Parainfluenza 2 | |
| Staphylococcus epidermidis | Parainfluenza 3 | |
| Streptococcus pneumoniae | Respiratory Syncytial virus, type B | |
| Streptococcus salivarius | Epstein Barr Virus | |
| Streptococcus pyogenes |
- Some cross-reactivity was observed for E. coli concentrations greater than 2.20 x 10°, Moraxella cat concentrations greater than 2.40 x 10°, and Proteus vulgaris at concentrations greater than 1.50 x 10°
In addition, in silico bioinformatics analyses were performed to assess the level of sequence similarity between the influenza A and B target nucleic acid sequence and the genomic sequences of the following upper respiratory tract microorganism. None of the organisms maintained genomic sequence that was significantly similar to the Alere™ i Influenza A & B 2 target sequences.
| Bacteria | Viruses |
|---|---|
| Bordetella bronchiseptica | Adenovirus 2 |
| Chlamydia pneumoniae | Adenovirus 3 |
| Chlamydia trachomatis | Adenovirus 4 |
| Neisseria gonorrhoae | Adenovirus 5 |
| Neisseria mucosa | Adenovirus 11 |
| Proteus mirabilis | Adenovirus 14 |
| Adenovirus 31 | |
| Coronavirus NL63 | |
| Coxsackievirus B35 | |
| Echovirus 6 | |
| Echovirus 9 | |
| Echovirus 11 |
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Enterovirus 71
INTERFERING SUBSTANCES
The following substances, naturally present or artificially introduced into the nasal cavity or nasopharynx were evaluated with Alere™ i Influenza A & B 2 at the concentrations listed below and were found not to affect test performance.
| Substance | Concentration |
|---|---|
| Mucin | 0.5% w/v |
| Whole Blood | 1% v/v |
| NeoSynephrine Nasal Spray | 20% v/v |
| Afrin Original Nasal Spray | 20% v/v |
| Ocean Saline Nasal Spray | 20% v/v |
| Chloroseptic Max | 20% w/v |
| Zicam | 20% v/v |
| Beclomethasone | 0.068 mg/mL |
| Budesonide | 0.051 mg/mL |
| Dexamethasone | 0.48 mg/mL |
| Flunisolide | 0.04 mg/mL |
| Fluticasone | 0.04 mg/mL |
| Mometasone | 0.04 mg/mL |
| Mupirocin | 4.3 mg/mL |
| Tobryamycin | 1.44 mg/mL |
| Triamcinolone | 0.04 mg/mL |
| Zanamivir (Relenza) | 0.284 mg/mL |
Inhibition by other Microorganisms
Alere™ i Influenza A & B test performance in the presence of non-influenza respiratory pathogens was evaluated. Vendor provided stocks of influenza A and B strains were diluted in UTM to approximately 3 times the limit of detection. Contrived influenza A and B positive swab specimens were prepared by coating 10 virus dilution onto each swab. The following panel of non-influenza viruses were tested at the concentration provided in the table below and was found not to affect test performance.
| Virus Panel | Concentration |
|---|---|
| Adenovirus Type 1 | 2.95 x 107 TCID50/mL |
| Rhinovirus Type 1A | 1.58 x 108 TCID50/mL |
| Respiratory Syncytial Virus, Type B, Strain 18537 | 3.00 x 103 (PFU/mL) |
Inhibition by High Levels of Influenza A and B
Alere™ i Influenza A & B test performance in the presence of influenza A and B was evaluated. Vendor provided stocks of influenza A and B strains were diluted in UTM to approximately 3 times the limit of detection. Contrived influenza A and B positive swab specimens were prepared by coating 10 microliters of virus dilution onto each swab. To create the co-infection swabs, diluted influenza A (at a concentration approximately 30 times the LoD) was added to the near LoD Flu B swab. Likewise, diluted influenza B (at a concentration approximately 30 times the LoD) was added to the near LoD Flu A swab. No impact on test performance was observed.
Carry-Over Contamination
An analytical carry-over study was performed to demonstrate that when recommended laboratory practices are followed, there is little risk of false positive results caused by carryover or cross-contamination in the Alere™ i Influenza A & B 2 test. Vendor provided stocks of influenza A and B strains were diluted in UTM to approximately 30 times the limit of detection. Contrived influenza A and B positive swab specimens were prepared by coating 10
{11}------------------------------------------------
microliters of virus dilution onto each swab. Testing of the contrived positive swabs was alternated with testing of a negative swab sample for a total of 15 rounds. There were no false positive results obtained.
An additional analytical carry-over study was performed testive VTM samples alternated with negative VTM samples following the test procedure for Nasal or Nasopharyngeal Swab Eluted in Viral Transport Media for a total of 15 rounds. There were no false positive results obtained.
REPRODUCIBILITY
A reproducibility study of Alere™ i Influenza A & B 2 was conducted by operators from 3 sites using panels of blind coded specimens containing negative (at the limit of detection), and moderate positive (above the limit of detection) influenza A and B samples. Participants tested each sample multiple times on 5 different days. The percent agreement with expected results for the influenza A moderate positive samples was 100% (90/90). The percent agreement with expected result for the influenza B moderate positive and low positive samples were 100% (89/89), 98.9% (89/90), respectfully. All of the true negative samples (89) generated negative test results. There were no significant differences observed within run (replicates tested by one operator), between run (five different days), between sites (three sites), or between operators (nine operators).
§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.
(a)
Identification. A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B;
(2) Influenza A subtype H1 and Influenza A subtype H3;
(3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B;
(4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus;
(5) Human Metapneumovirus;
(6) Rhinovirus; and
(7) Adenovirus.
(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;”
(2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and
(3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.