(51 days)
The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA RSV/MPV Identification Kit is intended for the qualitative identification of respiratory syncytial virus and human metapneumovirus in nasal and nasopharyngeal swabs and aspirates/washes specimens from patients with signs and symptoms of respiratory infection by direct detection of immunofluorescence using monoclonal antibodies (MAbs).
It is recommended that specimens found to be negative for respiratory syncytial virus after examination of the direct specimen result be confirmed by cell culture. Specimens found to be negative for human metapneumovirus after examination of the direct specimen results should be confirmed by an FDA-cleared human metapneumovirus molecular assay. Negative results do not preclude respiratory syncytial virus and human metapneumovirus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.
The D3 FastPoint L-DFA RSV/MPV Identification Kit uses a blend (called a "L-DFA Reagent'") of viral antigen-specific murine monoclonal antibodies that are directly labeled with either R-phycoerythin (PE) (respiratory syncytial virus) or fluorescein isothiocyanate (FITC) (human metapneumovirus) for the rapid identification of respiratory syncytial virus and human metapneumovirus in nasal and nasopharyngeal swabs and aspirates from patients with signs and symptoms of respiratory infection.
The cells to be tested are derived from respiratory specimens from patients with signs and symptoms of respiratory infection. The cells are permeabilized and stained concurrently in a liquid suspension format with the L-DFA Reagent. After incubating at 35℃ to 37℃ for 5-minutes, the stained cell suspensions are rinsed with 1X PBS. The rinsed cells are pelleted by centrifugation and then re-suspended with the Resuspension Buffer and loaded onto a specimen slide well. The cells are examined using a fluorescence microscope. Cells infected with RSV will exhibit golden-yellow fluorescence due to the PE. Cells infected with hMPV will exhibit apple-green fluorescence due to the FITC. Non-infected cells will exhibit red fluorescence due to the Evans Blue counter-stain. Nuclei of intact cells will exhibit orange-red fluorescence due to the propidium iodide.
Acceptance Criteria and Study for D3 FastPoint L-DFA RSV/MPV Identification Kit
1. Table of Acceptance Criteria and Reported Device Performance
The provided document doesn't explicitly state numerical acceptance criteria for all performance metrics. However, for a 510(k) submission, implied acceptance is often "substantially equivalent" to predicate devices, and for clinical performance, high sensitivity and specificity are expected. The reproducibility study explicitly aims for 100% agreement.
Note: The "acceptance criteria" are inferred based on standard expectations for diagnostic device performance and the detailed reporting of study outcomes, particularly the 100% agreement for reproducibility and the very high sensitivity/specificity for RSV. The lower sensitivity for hMPV in NP swab samples might be within acceptable limits given the context of medical device approval for challenging targets.
| Performance Metric | Acceptance Criteria (Implied/Explicit) | Reported Device Performance (D3 FastPoint L-DFA RSV/MPV Identification Kit) |
|---|---|---|
| Reproducibility (Overall Agreement with Expected Result) | 100% (Implied by study design expecting full agreement) | 100% (280/280) across all sites and panel members |
| Limit of Detection (LoD) - RSV | The lowest dilution at which at least 9/10 replicates are detected. | 100 infected cells/mL |
| Limit of Detection (LoD) - hMPV | The lowest dilution at which at least 9/10 replicates are detected. | 100 infected cells/mL |
| Analytical Reactivity (Inclusivity) - RSV | Detection of various RSV strains at 10x LoD. | All 3 tested RSV strains detected at 10x LoD. |
| Analytical Reactivity (Inclusivity) - hMPV | Detection of various hMPV strains at 10x LoD. | All 4 tested hMPV strains detected at 10x LoD. |
| Clinical Sensitivity (RSV - NP wash/aspirate) | High sensitivity for diagnosis. | 98.6% (204/207) [95% CI: 95.8-99.7%] |
| Clinical Specificity (RSV - NP wash/aspirate) | High specificity for diagnosis. | 99.8% (462/463) [95% CI: 98.8-100%] |
| Clinical Sensitivity (hMPV - NP wash/aspirate) | High sensitivity for diagnosis. | 68.8% (55/80) [95% CI: 57.4-78.7%] |
| Clinical Specificity (hMPV - NP wash/aspirate) | High specificity for diagnosis. | 100.0% (614/614) [95% CI: 99.4-100%] |
| Clinical Sensitivity (RSV - NP swab) | High sensitivity for diagnosis. | 97.5% (39/40) [95% CI: 86.8-99.9%] |
| Clinical Specificity (RSV - NP swab) | High specificity for diagnosis. | 100.0% (647/647) [95% CI: 99.4-100%] |
| Clinical Sensitivity (hMPV - NP swab) | High sensitivity for diagnosis. | 54.5% (24/44) [95% CI: 38.8-69.9%] |
| Clinical Specificity (hMPV - NP swab) | High specificity for diagnosis. | 100.0% (632/632) [95% CI: 99.4-100%] |
2. Sample Size Used for the Test Set and Data Provenance
- Reproducibility Test Set:
- Sample Size: A reproducibility panel consisting of 5 members (low RSV, low hMPV, mixed RSV/hMPV, mixed hMPV/RSV, negative). Each panel member was tested daily in two separate runs for 5 days by 4 different laboratories, resulting in 40 total runs. This yielded 280 total tests (across all panel members and runs) with individual results reported for expected positive and negative wells.
- Data Provenance: The study was conducted at four different laboratories. The document does not specify the country of origin but implies U.S. clinical laboratories (referencing "U.S. clinical laboratories" for clinical performance). It's a prospective study in the sense that the testing itself was performed to assess reproducibility.
- Limit of Detection (LoD) Test Set:
- Sample Size: Dilution series of infected model cells were used. For each virus (RSV and hMPV A1), 10 replicate microscope slides were prepared for each dilution level. The specific number of dilutions isn't explicitly stated as a single number but spanned from 1000 infected cells/mL down to 0.8 or 1.5 infected cells/mL, with 10 replicates for each dilution.
- Data Provenance: Laboratory study, likely internal to the manufacturer or a contracted lab.
- Analytical Reactivity (Inclusivity) Test Set:
- Sample Size: 3 RSV virus strains and 4 hMPV virus strains were evaluated. For each strain, "low concentration infected cell suspensions (approximately 4% cells infected, 25 to 50 infected cells)" were prepared.
- Data Provenance: Laboratory study.
- Clinical Performance Test Set:
- Sample Size: 1519 total respiratory specimens (nasal and nasopharyngeal swabs and aspirates/washes).
- Data Provenance: Prospective studies at 4 geographically diverse U.S. clinical laboratories during the 2009 respiratory virus seasons (January 2009 - March 2009). The specimens were "excess, remnants of respiratory specimens that were prospectively collected from symptomatic individuals suspected of respiratory infection, and were submitted for routine care or analysis by each site, and that otherwise would have been discarded." Individual specimens were delinked from patient identifiers.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications
The document does not explicitly state the number or qualifications of experts for establishing ground truth as a separate role. Instead, the ground truth for clinical performance was established using a composite comparator method:
- RSV: Direct Specimen Fluorescent Antibody (DSFA) test with an FDA-cleared predicate device, followed by viral culture confirmation of all negatives from the comparator DSFA test.
- hMPV: DSFA with an FDA-cleared predicate device, followed by confirmation of all negative specimens from the comparator DSFA test using a validated hMPV real-time RT-PCR, which was then followed by bi-directional sequencing analysis.
This implies that the "ground truth" was determined by the results of these established and confirmed laboratory methods, rather than by human expert consensus or adjudication of raw images/output from the test device solely.
4. Adjudication Method for the Test Set
For the clinical studies, an explicit "adjudication method" involving human experts reviewing conflicting results is not detailed. Instead, a composite comparator algorithm was used to define "true positive" and "true negative":
- "True positive" for RSV was defined as any sample that tested positive by the comparator DSFA test or viral culture.
- "True positive" for hMPV was defined as any sample that tested positive by the comparator DSFA test OR had bi-directional sequencing data meeting pre-defined quality acceptance criteria that matched hMPV sequences in GenBank.
- "True negative" was defined as any sample that tested negative by both the comparator DSFA test and either viral culture (for RSV) or the hMPV real-time RT-PCR comparator assay (for hMPV).
This approach essentially pre-defines how discordant results between screening and confirmatory tests contribute to the final ground truth, replacing a separate human adjudication step.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done
No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not conducted. This study is an evaluation of an in-vitro diagnostic device (IVD), specifically a direct fluorescent antibody (DFA) test, which is read by trained laboratory personnel, but the study focuses on the device's performance against comparator methods, not on comparing reader performance with and without AI assistance (as it is not an AI device).
6. If a Standalone (i.e. algorithm only without human-in-the loop performance) was done
Yes, the studies presented are essentially standalone performance evaluations of the device. The D3 FastPoint L-DFA RSV/MPV Identification Kit is an immunofluorescent assay where a human technician observes fluorescent staining patterns under a microscope. However, the performance metrics (sensitivity, specificity) are for the device's ability to detect the viral antigens in specimens, without involving a study design where human readers using the device are compared to human readers using another method, or AI assistance. The results in the tables reflect the diagnostic performance of the kit itself when used according to its instructions.
7. The Type of Ground Truth Used (expert consensus, pathology, outcomes data, etc)
The ground truth used for the clinical performance studies was based on a composite comparator method combining:
- FDA-cleared predicate DFA devices
- Viral culture (for RSV)
- Validated real-time RT-PCR with bi-directional sequencing analysis (for hMPV)
This is a form of reference standard composite, aiming for a highly accurate and confirmed diagnosis of the presence or absence of the target viruses.
8. The Sample Size for the Training Set
The document does not explicitly describe a separate "training set" or "validation set" in the context of machine learning. This is a traditional in vitro diagnostic device, not an AI/ML-based device.
All the described studies (reproducibility, LoD, analytical reactivity, clinical performance) contribute to the overall evidence for the device. The 1519 clinical specimens (fresh nasal/nasopharyngeal wash/aspirate and swab specimens) can be considered the test set for evaluating clinical performance.
9. How the Ground Truth for the Training Set Was Established
As noted above, there is no explicit "training set" mentioned in the context of machine learning. The studies assess the performance of the pre-developed D3 FastPoint L-DFA RSV/MPV Identification Kit using the specified ground truth methods.
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Diagnostic Hybrids, Inc.
D3 FastPoint L-DFA RSV/MPV Identification Kit
10/06/2009 Page 1 of 14
Section 05, 510(k) Summary
Applicant:
DIAGNOSTIC HYBRIDS, INC. 1055 East State Street Suite 100 Athens, OHIO 45701
Contact Information:
Ronald H. Lollar, Senior Director Product Realization, Management, and Marketing 1055 East State Street Suite 100 Athens, Ohio 45701 740-589-3300 - Corporate number 740-589-3373 - Desk phone 740-593-8437 -- Fax lollar@dhiusa.com
Date of preparation of 510(k) summary:
October 5. 2009
Device Name:
Trade name - D FastPoint L-DFA RSV/MPV Identification Kit Common name - RSV/MPV DFA assay Classification name - Respiratory viral panel multiplex nucleic acid assay Product Code - OMG, LKT Regulation - 21 CFR 866.3980 Regulatory Class - Class II Panel Microbiology (83)
Legally marketed devices to which equivalence is claimed:
D3 Ultra DFA Respiratory Virus Screening & ID Kit (K061101)
Intended Use: The Diagnostic Hybrids, Inc. D3 Ultra DFA (direct fluorescent antibody) Respiratory Virus Screening & ID Kit (D3 Ultra) is intended for the qualitative detection and identification of the influenza A, influenza B, respiratory syncytial virus (RSV), adenovirus, parainfluenza 1, parainfluenza 2 and parainfluenza 3 virus in respiratory specimens, by either
DEC - 4 2009
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direct detection or cell culture method, by immunofluorescence using fluoresceinated monoclonal antibodies (MAbs). It is recommended that specimens found to be negative after examination of the direct specimen result be confirmed by cell culture. Negative results do not preclude respiratory virus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.
- Performance characteristics for influenza A were established when . influenza A/H3 and A/H1 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 departments for testing. Viral culture should not be attempted in these cases unless a BSL3+ facility is available to receive and culture specimens.
D' Duet DFA RSV/Respiratory Virus Screening Kit (K081928)
Intended Use: The Diagnostic Hybrids, Inc. device, D3 Duet DFA RSV/Respiratory Virus Screening Kit (D3 Duet RSV Kit), is intended for the qualitative detection and identification of respiratory syncytial virus, while screening for influenza A virus, influenza B virus, adenovirus, and parainfluenza virus types 1, 2 and 3 viral antigens, in nasal and nasopharyngeal swabs and aspirates or in cell culture. The assay detects viral antigens by immunofluorescence using monoclonal antibodies (MAbs), from patients with signs and symptoms of respiratory infection.
It is recommended that specimens found to be negative after examination of the direct specimen result be confirmed by cell culture. Negative results do not preclude influenza virus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.
Performance characteristics for influenza A virus detection and identification were established when influenza A (H3N2) and influenza A (H1N1) were the predominant influenza A strains circulating in the United States. Performance characteristics for influenza A virus detection and identification were established when influenza A H3N2 and influenza A H1N1 were the predominant influenza A strains circulating in the United States. 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
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recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to a state or local health department for testing. Viral culture should not be attempted in these cases unless a BSL3+ facility is available to receive and culture specimens.
D3 DFA Metapneumovirus Identification Kit (K090073)
Intended Use: The Diagnostic Hybrids, Inc. device, D3 DFA Metapneumovirus Identification Kit (D3 MPV Kit), is intended for the qualitative detection and identification of human metapneumovirus (hMPV) in nasal and nasopharyngeal swabs and aspirates/washes or cell culture. The assay detects hMPV antigens by immunofluorescence using a blend of three monoclonal antibodies (MAbs), from patients with signs and symptoms of acute respiratory infection. This assay detects but is not intended to differentiate the four recognized genetic sub-lineages of hMPV.
Negative results do not preclude hMPV infection and should not be used as the sole basis for diagnosis, treatment or other management decisions. It is recommended that specimens found to be negative after examination of the direct specimen results be confirmed by an FDA-cleared hMPV molecular assay.
Device Description:
The D3 FastPoint L-DFA RSV/MPV Identification Kit uses a blend (called a "L-DFA Reagent'") of viral antigen-specific murine monoclonal antibodies that are directly labeled with either R-phycoerythin (PE) (respiratory syncytial virus) or fluorescein isothiocyanate (FITC) (human metapneumovirus) for the rapid identification of respiratory syncytial virus and human metapneumovirus in nasal and nasopharyngeal swabs and aspirates from patients with signs and symptoms of respiratory infection.
Kit Components:
- D3 FastPoint L-DFA RSV/MPV Reagent, 4.0-mL. One dropper bottle 1. containing a mixture of PE-labeled murine monoclonal antibodies directed against respiratory syncytial virus antigens and FITC-labeled murine monoclonal antibodies directed against human metapneumovirus antigens. The buffered, stabilized, aqueous solution contains Evans Blue and propidium iodide as counter-stains and 0.1% sodium azide as preservative.
- 40X PBS Concentrate, 25-mL. One bottle of 40X PBS concentrate containing 2. 4% sodium azide (0.1% sodium azide after dilution to 1X using de-mineralized water).
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- Re-suspension Buffer, 6.0-mL. One bottle of a buffered glycerol solution and 3. 0.1% sodium azide.
- D3 FastPoint L-DFA RSV/MPV Antigen Control Slides, 5-slides. Five 4. individually packaged control slides containing 2 wells with cell culture-derived positive and negative control cells. Each positive well contains cells infected with either respiratory syncytial virus or human metapneumovirus. The negative wells contain non-infected cells. Each slide is intended to be stained only one time.
- D3 FastPoint L-DFA Specimen Slides and Coverslips, 50-slides with 5. coverslips. Fifty pack of 3-well specimen slides.
The cells to be tested are derived from respiratory specimens from patients with signs and symptoms of respiratory infection. The cells are permeabilized and stained concurrently in a liquid suspension format with the L-DFA Reagent. After incubating at 35℃ to 37℃ for 5-minutes, the stained cell suspensions are rinsed with 1X PBS. The rinsed cells are pelleted by centrifugation and then re-suspended with the Resuspension Buffer and loaded onto a specimen slide well. The cells are examined using a fluorescence microscope. Cells infected with RSV will exhibit golden-yellow fluorescence due to the PE. Cells infected with hMPV will exhibit apple-green fluorescence due to the FITC. Non-infected cells will exhibit red fluorescence due to the Evans Blue counter-stain. Nuclei of intact cells will exhibit orange-red fluorescence due to the propidium iodide.
Intended Use:
The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA RSV/MPV Identification Kit is intended for the qualitative identification of respiratory syncytial virus and human metapneumovirus in nasal and nasopharyngeal swabs and aspirates/washes specimens from patients with signs and symptoms of respiratory infection by direct detection of immunofluorescence using monoclonal antibodies (MAbs).
It is recommended that specimens found to be negative for respiratory syncytial virus after examination of the direct specimen result be confirmed by cell culture. Specimens found to be negative for human metapneumovirus after examination of the direct specimen results should be confirmed by an FDA-cleared human metapneumovirus molecular assay. Negative results do not preclude respiratory syncytial virus and human metapneumovirus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions. ·
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| TABLE 5.1: Characteristics of the D3 FastPoint L-DFA Kit are compared to those of the followingDiagnostic Hybrids (DHI) predicate devices | |||||
|---|---|---|---|---|---|
| Characteristics | D3 FastPointRSV/MPVIdentification Kit(Subject Device) | D3 Ultra Kit510(k) #K061101 | D3 Duet RSV Kit510(k) # K081928 | D3 MPV Kit510(k) # K090073 | |
| Intended Use | The DiagnosticHybrids, Inc. device,D3 FastPoint L-DFARSV/MPVIdentification Kit isintended for thequalitativeidentification ofrespiratory syncytialvirus and humanmetapneumovirus innasal andnasopharyngealswabs andaspirates/washesspecimens frompatients with signsand symptoms ofrespiratory infectionby direct detectionofimmunofluorescenceusing monoclonalantibodies (MAbs).It is recommendedthat specimensfound to be negativefor respiratorysyncytial virus afterexamination of thedirect specimenresult be confirmedby cell culture.Specimens found tobe negative forhumanmetapneumovirusafter examination ofthe direct specimenresults should beconfirmed by anFDA-cleared humanmetapneumovirusmolecular assay.Negative results do | The DiagnosticHybrids, Inc. D3UltraTM DFA (directfluorescentantibody)Respiratory VirusScreening & ID Kitis intended for thequalitative detectionand identification ofthe influenza A,influenza B,respiratory syncytialvirus (RSV),adenovirus,parainfluenza 1,parainfluenza 2 andparainfluenza 3virus in respiratoryspecimens, by eitherdirect detection orcell culture method,byimmunofluorescenceusing monoclonalantibodies (MAbs).It is recommendedthat specimensfound to be negativeafter examination ofthe direct specimenresult be confirmedby cell culture.Negative results donot precluderespiratory virusinfection and shouldnot be used as thesole basis fordiagnosis, treatmentor othermanagementdecisions. | The DiagnosticHybrids, Inc. device,D3 Duet DFARSV/RespiratoryVirus Screening Kit,is intended for thequalitative detectionand identification ofrespiratory syncytialvirus, whilescreening forinfluenza A virus,influenza B virus,adenovirus, andparainfluenza virustypes 1, 2 and 3 viralantigens, in nasaland nasopharyngealswabs and aspiratesor in cell culture.The assay detectsviral antigens byimmunofluorescenceusing monoclonalantibodies (MAbs),from patients withsigns and symptomsof respiratoryinfection.It is recommendedthat specimensfound to be negativeafter examination ofthe direct specimenresult be confirmedby cell culture.Negative results donot precludeinfluenza virus .infection and shouldnot be used as thesole basis fordiagnosis, treatmentor othermanagement | The DiagnosticHybrids, Inc. device,D3 DFAMetapneumovirusIdentification Kit, isintended for thequalitative detectionand identification ofhumanmetapneumovirus(hMPV) in nasal andnasopharyngealswabs andaspirates/washes orcell culture. Theassay detects hMPVantigens byimmunofluorescenceusing a blend ofthree monoclonalantibodies (MAbs),from patients withsigns and symptomsof acute respiratoryinfection. Thisassay detects but isnot intended todifferentiate the fourrecognized geneticsub-lineages ofhMPV.Negative results donot preclude hMPVinfection and shouldnot be used as thesole basis fordiagnosis, treatmentor othermanagementdecisions.It is recommendedthat specimens foundto be negative afterexamination of thedirect specimen | |
| TABLE 5.1: Characteristics of the D3 FastPoint L-DFA Kit are compared to those of the followingDiagnostic Hybrids (DHI) predicate devices | |||||
| Characteristics | D3 FastPointRSV/MPVIdentification Kit(Subject Device) | D3 Ultra Kit510(k) #K061101 | D3 Duet RSV Kit510(k) # K081928 | D3 MPV Kit510(k) # K090073 | |
| not precluderespiratory virusinfection and shouldnot be used as thesole basis fordiagnosis, treatmentor othermanagementdecisions. | decisions. | results be confirmedby an FDA-clearedhMPV molecularassay. | |||
| Target Viruses | respiratory syncytialvirus,metapneumovirus | influenza A virus,influenza B virus,respiratorysyncytial virus,adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 3 | influenza A virus,influenza B virus,respiratorysyncytial virus,adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 3 | metapneumovirus | |
| Monoclonal antibodies(MAbs) | The D3 FastPoint L-DFA RSV/MPVReagent contain 5MAbs to respiratorysyncytial virus (2)andmetapneumovirus(3) | The RespiratoryVirus DFAScreening Reagentcontains 15 MAbs to7 differentrespiratory viruses(influenza A virus,influenza B virus,respiratory syncytialvirus, adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 3) | TheRSV/RespiratoryVirus DFAScreening Reagentcontains 15 MAbs to7 differentrespiratory viruses(influenza A virus,influenza B virus,adenovirus,parainfluenza virustype 1, parainfluenzavirus type 2,parainfluenza virustype 3), plus 2 MAbsto respiratorysyncytial virus. | TheMetapneumovirusDFA Reagentcontains 3 MAbs tometapneumovirus | |
| Labeling method | Direct labeling- using R-Phycoerythrin (R-PE) to label theMAbs to RSV. | Direct labeling- using fluoresceinisothiocyanate(FITC) to label allMAbs withfluorescein. | Direct labeling- using R-Phycoerythrin (R-PE) to label theMAbs to respiratorysyncytial virus.- using fluoresceinisothiocyanate | Direct labeling- using fluoresceinisothiocyanate(FITC) to label allMAbs withfluorescein. | |
| TABLE 5.1: Characteristics of the D³ FastPoint L-DFA Kit are compared to those of the followingDiagnostic Hybrids (DHI) predicate devices | |||||
| Characteristics | D³ FastPointRSV/MPVIdentification Kit(Subject Device) | D³ Ultra Kit510(k) #K061101 | D³ Duet RSV Kit510(k) #K081928 | D³ MPV Kit510(k) # K090073 | |
| (FITC) to label theMAbs tometapneumovirus. | (FITC) to label allother MAbs withfluorescein. | ||||
| R-Phycoerythrin-labeledMAbs | respiratory syncytialvirus | None | respiratory syncytialvirus | None | |
| Fluorescein-labeled MAbs | metapneumovirus | influenza A virus,influenza B virus,respiratory syncytialvirus, adenovirus,parainfluenzavirustype 1,parainfluenzavirustype 2,parainfluenzavirustype 3 | influenza A virus,influenza B virus,adenovirus,parainfluenzavirustype 1, parainfluenzavirus type 2,parainfluenzavirustype 3 | metapneumovirus | |
| Cell Fixative | Proprietary Non-Acetone basedsystem | Acetone | Acetone | Acetone | |
| Cell Counter-stain | Propidium Iodide,Evans Blue | Evans Blue | Evans Blue | Evans Blue | |
| Performance characteristics | |||||
| Staining patterns | RespiratorySyncytial Virus:The fluorescence iscytoplasmic. Cellsappear round.Metapneumovirus:The fluorescence iscytoplasmic andpunctate. Cellsappear round.Negative: Cellsfluoresce red due tothe Evans Bluecounter-stain.Nuclei: Cell Nucleifluoresce orange-reddue to thePropidium Iodidecounter-stain. | Influenza A and B:The fluorescence iscytoplasmic, nuclearor both.Cytoplasmicstaining is oftenpunctate with largeinclusions whilenuclear staining isuniformly bright.RespiratorySyncytial Virus:The fluorescence iscytoplasmic andpunctate with smallinclusions in thesyncytia.Parainfluenza 1, 2,3: The fluorescenceis cytoplasmic andpunctate withirregular inclusions.Types 2 and 3 causethe formation ofsyncytia. | Influenza A andB: Thefluorescence iscytoplasmic,nuclear or both.Cytoplasmicstaining is oftenpunctate with largeinclusions whilenuclear staining isuniformly bright.RespiratorySyncytial Virus:The fluorescenceis cytoplasmic andpunctate withsmall inclusions inthe syncytia.Parainfluenza 1,2, 3: Thefluorescence iscytoplasmic andpunctate withirregularinclusions. Types | Metapneumovirus:The fluorescence iscytoplasmic andpunctate with smallinclusions in thesyncytia.Negative: Entirecell fluoresce reddue to the EvansBlue counter-stain. | |
| TABLE 5.1: Characteristics of the D³ FastPoint L-DFA Kit are compared to those of the followingDiagnostic Hybrids (DHI) predicate devices | |||||
| Characteristics | D³ FastPointRSV/MPVIdentification Kit(Subject Device) | D³ Ultra Kit510(k) #K061101 | D³ Duet RSV Kit510(k) # K081928 | D³ MPV Kit510(k) # K090073 | |
| fluorescence iscytoplasmic andpunctate or brightnuclear or both.Negative: Cellsfluoresce red due tothe Evans Bluecounter-stain. | formation ofsyncytia.Adenovirus: Thefluorescence iscytoplasmic andpunctate or brightnuclear or both.Negative: Cellsfluoresce red dueto the Evans Bluecounter-stain. | ||||
| Device Reagents are not reactive with these numbers of microorganisms. | |||||
| Analyticalspecificity(cross-reactivitystudies; variousstrains ofmicroorganismsand cell lines) | Viruses | 59 | 31 | 32 | 59 |
| Bacteria | 22 | 18 | 25 | 25 | |
| Chlamydiaspp. | 1 | 1 | 3 | 3 | |
| Yeast | 1 | 0 | 1 | 1 | |
| Protozoan | 0 | 0 | 1 | 1 | |
| Cell lines | N/A | 17 | 17 | 16 |
Technological Characteristics, Compared to Predicate Device:
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Analytical Performance:
Precision/Reproducibility:
Assay precision, intra-assay variability and inter assay variability were assessed with a reproducibility panel consisting of 5 randomized panel members.
The RSV/hMPV panel consisted of the following:
- a. Low level RSV (Washington strain) infected cells.
- b. Low level hMPV (A1 subtype) infected cells.
- c. Low level RSV (Washington strain) infected cells mixed with mid level hMPV (A1 subtype) infected cells.
- d. Low level hMPV (A1 subtype) infected cells mixed with mid level RSV (Washington strain) infected cells.
- e. Mid level non-infected (negative) cells.
The low level is estimated to contain between 4 to 10% infected cells in the sample. The mid level is estimated to contain between 20 to 25% infected cells in the sample. Each sample contains 2.5 x106 to 3.5 x105 total cells.
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Each panel was tested daily in two separate runs for 5-days by four different laboratories (40 total runs). The following results were recorded:
- a. Presence or absence of golden-yellow fluorescence.
- b. Percent of cells exhibiting golden-yellow fluorescence.
- c. Presence or absence of apple-green fluorescence.
- d. Percent of cells exhibiting apple-green fluorescence.
For the L-DFA Reagent, the combined data from the four Study Sites demonstrated reproducible detection of RSV by the R-PE labeled MAbs and reproducible detection of hMPV by the FITC-labeled MAbs. The presence of RSV infected cells was reported in 100% (120/120) of the wells in which the infected cells were expected. The presence of hMPV infected cells was reported in 100% (120/120) of the wells in which the infected cells were expected. The absence of infected cells was reported in 100% (40/40) of the wells in which infected cells were not present. The total percent agreement for the L-DFA Reagent was 100% (280/280):
| TABLE 5.2: Reproducibilty Study Results using the L-DFA Reagent | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Site | PanelMember | Negative | RSVLowLevel | hMPVLowLevel | Mixed Infection | Mixed Infection | |||
| RSV MidLevel | hMPVLow Level | RSVLowLevel | hMPVMidLevel | Total %Agreement | |||||
| Concentration | Noinfectedcells | 4 to 10%infectedcells | 4 to 10%infectedcells | 20 to30%infectedcells | 4 to 10%infectedcells | 4 to 10%infectedcells | 20 to30%infectedcells | ||
| Site1 | AgreementwithExpected result | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 70/70(100%) |
| Site2 | AgreementwithExpected result | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 70/70(100%) |
| Site3 | AgreementwithExpected result | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 70/70(100%) |
| Site4 | AgreementwithExpected result | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 70/70(100%) |
| Total Agreement withExpectedresult | 40/40(100%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 280/280(100%) | |
| 95% CI | 91.2 -100% | 91.2 -100% | 91.2 -100% | 91.2 -100% | 91.2 -100% | 91.2 -100% | 91.2 -100% | 98.7 - 100% |
Limit of Detection:
Analytical Limit of Detections (LoDs) of the L-DFA Reagent was addressed using dilution series of infected model cells. Model cells for respiratory syncytial virus (ATCC Washington strain) and human metapneumovirus
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10/06/2009
subtype A1 (clinical strain) were diluted with non-infected cells to produce a suspension equivalent to 1,000 infected cells per milliliter. This level theoretically yields approximately 25 infected cells per 25-uL of suspension. This suspension was then serially diluted to a theoretical level of less than 1 cell per milliliter. (NOTE: This level was the target to begin with a low positive level. Actual starting levels vary, however, and are within 1 dilution of the 25 infected cell target level). 25-uL aliquots from each dilution level were spotted onto 10 replicate microscope slides, and then stained according to the instructions for use described in this product insert. Each cell spot was examined at 200x magnification. Results were reported as numbers of positive replicates for each set of 10. Analytical detection limits for each of the 8 analytes were defined as the lowest dilutions at which at least 9 out of 10 replicates were detected. LoD study results are summarized in TABLE 5.3 below:
| TABLE 5.3: Limit of Detections of the L-DFA Reagent | |||
|---|---|---|---|
| Virus Strain | Infected cells/mL | Number of replicates with positive cells | LOD determination |
| RSV(ATCC Washingtonstrain) | 1000 | 10/10 | 100 infected cells/mL |
| 200 | 10/10 | ||
| 100 | 10/10 | ||
| 50 | 7/10 | ||
| 25 | 7/10 | ||
| 12.5 | 6/10 | ||
| 6 | 1/10 | ||
| 3 | 0/10 | ||
| 1.5 | 0/10 | ||
| 0.8 | 0/10 | ||
| hMPV A1(Clinical strain) | 2000 | 10/10 | 100 infected cells/mL |
| 400 | 10/10 | ||
| 200 | 10/10 | ||
| 100 | 10/10 | ||
| 50 | 6/10 | ||
| 25 | 2/10 | ||
| 12.5 | 0/10 | ||
| 6 | 0/10 | ||
| 3 | 0/10 | ||
| 1.5 | 0/10 |
Analytical reactivity (inclusivity):
Analytical reactivity (inclusivity) of the L-DFA Reagent was evaluated using 3-RSV virus and 4-hMPV virus strains. Low concentration infected cell suspensions (approximately 4% cells infected, 25 to 50 infected cells) were prepared for each viral strain. The suspensions were stained with the L-DFA Reagent.
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| TABLE 5.4: Analytical Reactivity (inclusivity) of the L-DFA Reagent on various RSV virus and hMPV virus strains | ||
|---|---|---|
| RSV and hMPV Strains | Infected Cell Concentration(as multiples of the respectiveestablished LoD concentration) | L-DFA Reagent Results |
| RSV 9320 | 10x LoD | 22 Golden-yellow fluorescent cells |
| RSV Washington | 10x LoD | 22 Golden-yellow fluorescent cells |
| RSV Long | 10x LoD | 32 Golden-yellow fluorescent cells |
| hMPV A1 | 10x LoD | 25 Apple-green fluorescent cells |
| hMPV A2 | 10x LoD | 25 Apple-green fluorescent cells |
| hMPV B1 | 10x LoD | 25 Apple-green fluorescent cells |
| hMPV B2 | 10x LoD | 37 Apple-green fluorescent cells |
Clinical Performance:
Performance of the Dr FastPoint RSV/MPV Kit testing direct respiratory specimens were established during prospective studies at 4 geographically diverse U.S. clinical laboratories during the 2009 respiratory virus seasons (January 2009 - March 2009). All specimens used in the studies meeting the inclusion and exclusion criteria represented excess, remnants of respiratory specimens that were prospectively collected from symptomatic individuals suspected of respiratory infection, and were submitted for routine care or analysis by each site, and that otherwise would have been discarded. Individual specimens were delinked from all patient identifiers and given a study sample code. All clinical sites were granted waivers of informed consent by their IRBs for this study.
Performance of the D3 FastPoint RSV/MPV Kit was assessed and compared to a predetermined algorithm that used composite comparator methods. The composite comparator methods for respiratory syncytial virus consisted of Direct Specimen Fluorescent Antibody (DSFA) test with an FDA-cleared device and viral culture confirmation of all the negatives (as determined by the comparator DSFA test). For human metapneumovirus the composite comparator methods consisted of DSFA with an FDA-cleared device, and confirmation of all negative specimens (as determined by the comparator DSFA test) using a validated ' hMPV real-time RT-PCR followed by bi-directional sequencing analysis comparator assay. The hMPV real-time RT-PCR comparator assay targets the hMPV Nucleocapsid gene. "True"
Sec05 FastPoint RSV MPV 09-10061 FDA.doc
Analytical validation of the real-time hMPV RT-PCR followed by bi-directional sequencing analysis comparator assay included analytical sensitivity and reactivity study, analytical specificity study, and extraction efficiency study. The analytical sensitivity (limit of detection or LoD) of the real-time hMPV RT-PCR followed by bi-directional sequencing analysis comparator assay was determined using quantified (TCIDs /mL) stocks of the 4 hMPV (subtypes A1, A2, B1 and B2) strains diluted in hMPV negative nasopharyngeal clinical matrix, and ranged from 10 - 50 TCID30mL.
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positive was defined as any sample that either tested positive by the comparator DSFA test or viral culture, or had bi-directional sequencing data meeting pre-defined quality acceptance criteria that matched hMPV sequences deposited in the National Center for Biotechnology Information (NCBI) GenBank database (www.ncbi.nlm.nih.gov), with acceptable E-values.2 "True" negative was defined as any sample that tested negative by both the comparator DSFA test and either viral culture or the hMPV real-time RT-PCR comparator assay.
Prevalence of RSV and hMPV within this population as determined by the D3 FastPoint RSV/MPV Kit direct specimen testing is noted in TABLE 5.5 below:
| TABLE 5.5: RSV/hMPV Prevalence* | |||
|---|---|---|---|
| Age | TotalSpecimensEvaluated | RSV# positive(prevalence) | hMPV# positive(prevalence) |
| 0 - 1 month | 55 | 15 (27.3%) | 2 (3.6%) |
| > 1 month to 2 years | 577 | 154 (26.7%) | 41 (7.1%) |
| > 2 years to 12 years | 391 | 25 (6.4%) | 17 (4.3%) |
| > 12 years to 21 years | 173 | 4 (2.3%) | 3 (1.7%) |
| 22 years to 30 years | 57 | 0 | 1 (1.8%) |
| 31 years to 40 years | 71 | 1 (1.4%) | 3 (4.2%) |
| 41 years to 50 years | 52 | 0 | 1 (1.9%) |
| 51 years to 60 years | 46 | 1 (2.2%) | 3 (6.5%) |
| 61 years to 70 years | 33 | 1 (3.0%) | 1 (3.0%) |
| 71 years to 80 years | 16 | 1 (6.3%) | 4 (25.0%) |
| 81 years and above | 7 | 1 (14.3%) | 0 |
| Age Not Reported | 41 | 0 | 1 (2.4%) |
| Total | 1519 | 203 (13.4%) | 77 (5.1%) |
| * There were 2 - respiratory syncytial virus + metapneumovirus co- | |||
| infections detected. |
TABLES 5.6 and 5.7 below show the study results of the NP wash/aspirate specimen type (Sites 1, 2, and 3 combined):
(http://www.ncbi.nlm.nih.gov/books/bv.fcgi?rid=handbook.section.614).
Sec05 FastPoint_RSV_MPV_09-10061 FDA.doc
2 The E-values generated from the clinical trials range from a low of 5e-78 to a high of 1e-20. The E-Value from NCBI BLAST Alignment indicates the statistical significance of a given pair-wise alignment and reflects the size of the database and the scoring system used. The lower the E-Value, the more significant the hit. A sequence alignment that has an E-Value of 1e-3 means that this similarity has a 1 in 1000 chance of occurring by chance alone.
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:
፡ ・
· , --- -
| TABLE 5.6: Respiratory syncytial virus | |||
|---|---|---|---|
| Fresh nasal/nasopharyngealwash/aspirate | Comparator DSFA(negatives followed by culture with DFA) | ||
| DHI DSFA | Positive | Negative | Total |
| Positive | 204 | 1 | 205 |
| Negative | 3 | 462 | 465 |
| Total | 207 | 463 | 670 |
| 95% CI | |||
| Sensitivity | 204/207 | 98.6% | 95.8-99.7% |
| Specificity | 462/463 | 99.8% | 98.8-100% |
| TABLE 5.7: Human.metapneumovirus | |||
|---|---|---|---|
| Fresh nasal/nasopharyngeal. wash/aspirate | Comparator DSFA(negatives confirmed by a validated hMPVreal-time RT-PCR followed by bi-directionalsequencing analysis comparator assay) | ||
| DHI DSFA | Positive | Negative | . Total |
| Positive | 55 | 0 | 55 |
| Negative | 25 | 614 | 639 |
| Total | 80 | 614 | 694 |
| 95% CI | |||
| Sensitivity | 55/80 | 68.8% | 57.4-78.7% |
| Specificity | 614/614 | 100.0% | 99.4-100% |
TABLES 5.8 and 5.9 below show the study results of the NP swab specimen type (Sites 3 and 4 combined):
| TABLE 5.8: Respiratory syncytial virus | |||
|---|---|---|---|
| Fresh nasal/nasopharyngealswab | Comparator DSFA(negatives followed by culture with DFA) | ||
| DHI DSFA | Positive | Negative | Total |
| Positive | 39 | 0 | 39 |
| Negative | 1 | 647 | 648 |
| Total | 40 | 647 | 687 |
| 95% CI | |||
| Sensitivity | 39/40 | 97.5% | 86.8-99.9% |
| Specificity | 647/647 | 100.0% | 99.4-100% |
.
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| TABLE 5.9: Human metapneumovirus | |||
|---|---|---|---|
| Fresh nasal/nasopharyngealswab | Comparator DSFA(negatives confirmed by a validated hMPVreal-time RT-PCR followed by bi-directionalsequencing analysis comparator assay) | ||
| DHI DSFA | Positive | Negative | Total |
| Positive | 24 | 0 | 24 |
| Negative | 20 | 632 | 652 |
| Total | 44 | 632 | 676 |
| 95% CI | |||
| Sensitivity | 24/44 | 54.5% | 38.8-69.9% |
| Specificity | 632/632 | 100.0% | 99.4-100% |
Overall at the four Study Sites, the performance results of the D3 FastPoint L-DFA RSV/MPV Identification Kit, when compared to those of the comparator devices, D3 Ultra DFA Respiratory Virus Screening & ID Kit, D3 Duet DFA RSV/Respiratory Virus Screening Kit and D3 DFA Metapneumovirus Identification Kit demonstrate that the devices detect RSV and hMPV antigens in a similar manner.
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Image /page/14/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized eagle with three stripes forming its body and wings. The eagle is facing right. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the eagle.
DEPARTMENT OF HEALTH & HUMAN SERVICES '
Public Health Service
Food and Drug Administration 10903 New Hampshire Avenue Document Mail Center-WO66-G609 Silver Spring, MD 20993-0002
Mr. Ronald H. Lollar Senior Director Product Realization, Management and Marketing Diagnostic Hybrids, Inc 1055 East State Street, Suite 100 Athens, Ohio 45701
DEC - 4 2009
Re: K093233
Trade/Device Name: D FastPoint L- DFA RSV/MPV Identification Kit Regulation Number: 21 CFR § 866.3980 Regulation Name: Respiratory viral antigens (respiratory syncytial virus and human metapneumovirus) Regulatory Class: II Product Code: OMG, LKT Dated: October 5, 2009 Received: October 14, 2009
Dear Mr. Lollar:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing 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 such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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
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CFR Part 807): labeling (21 CFR Parts 801 and 809); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).
This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.
If you desire specific information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5460. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). 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) 443-6597 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html
Sincerely yours.
Ula Schla fu
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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510(k) Number (if known): K093233
Device Name: D3 FastPoint L-DFA RSV/MPV Identification Kit
Indication for Use:
The Diagnostic Hybrids, Inc. device, D FastPoint L-DFA RSV/MPV Identification Kit is intended for the qualitative identification of respiratory syncytial virus and human metapneumovirus in nasal and nasopharyngeal swabs and aspirates/washes specimens from patients with signs and symptoms of respiratory infection by direct detection of immunofluorescence using monoclonal antibodies (MAbs).
It is recommended that specimens found to be negative for respiratory syncytial virus after examination of the direct specimen result be confirmed by cell culture. Specimens found to be negative for human metapneumovirus after examination of the direct specimen results should be confirmed by an FDA-cleared human metapneumovirus molecular assay. Negative results do not preclude respiratory syncytial virus and human metapneumovirus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.
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 IF NEEDED
Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)
Uve Schif
Division Sign-Off
Office of In Vitro Diagnostic Device Evaluation and Safety
510(k) k 093233
§ 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.