(142 days)
The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA Respiratory Virus Identification Kit is intended for the qualitative identification of influenza A virus, influenza B virus, respiratory syncytial virus, human metapneumovirus, adenovirus and to screen for the presence of parainfluenza virus types 1, 2, and 3 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 influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus or parainfluenza viruses 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 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. Since influenza strains display antigenic drift and shift from year to year, performance characteristics may vary. If infection with a novel influenza A virus is suspected, based on clinical and epidemiological screening criteria communicated by public health authorities, collect specimens following appropriate infection control precautions and submit to state or local health departments, for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
The D3 FastPoint L-DFA Respiratory Virus Identification Kit uses three blends (each called a "L-DFA Reagent") of viral antigen-specific murine monoclonal antibodies that are directly labeled with either R-PE (influenza A virus, respiratory syncytial virus, and parainfluenza virus) or fluorescein (influenza B virus, metapneumovirus, and adenovirus) for the rapid identification of respiratory viruses in nasal and nasopharyngeal swabs and aspirates from patients with signs and symptoms of respiratory infection.
Kit Components:
- D3 FastPoint L-DFA Influenza A/Influenza B Reagent, 4.0-mL. One dropper bottle containing a mixture of PE-labeled murine monoclonal antibodies directed against influenza A virus antigens and FITC-labeled murine monoclonal antibodies directed against influenza B virus antigens. The buffered, stabilized, aqueous solution contains Evans Blue and propidium iodide as counter-stains and 0.1% sodium azide as preservative.
- D3 FastPoint L-DFA RSV/MPV Reagent, 4.0-mL. One dropper bottle containing a mixture of PE-labeled murine monoclonal antibodies directed against respiratory syncytial virus antigens and FITC-labeled murine monoclonal antibodies directed against metapneumovirus antigens. The buffered, stabilized, aqueous solution contains Evans Blue and propidium iodide as counter-stains and 0.1% sodium azide as preservative.
- D3 FastPoint L-DFA PIV/Adenovirus Reagent, 4.0-mL. One dropper bottle containing a mixture of PE-labeled murine monoclonal antibodies directed against parainfluenza virus types 1, 2, or 3 antigens and FITClabeled murine monoclonal antibodies directed against adenovirus 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 4% sodium azide (0.1% sodium azide after dilution to 1X using de-mineralized water).
- Re-suspension Buffer, 6.0-mL. One bottle of a buffered glycerol solution and 0.1% sodium azide.
- D3 FastPoint L-DFA Respiratory Virus Antigen Control Slides, 5-slides. Five individually packaged control slides containing 6 wells with cell culture-derived positive and negative control cells. Each positive well is identified as to the virus infected cells present, i.e., influenza A virus, influenza B virus, respiratory syncytial virus, metapneumovirus, parainfluenza virus, and adenovirus. The negative wells contain noninfected cells. Each slide is intended to be stained only one time.
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 in 3 separate vials, each containing one of the 3 above reagents. 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 influenza A virus, respiratory syncytial virus, or parainfluenza virus types 1, 2 and 3 will exhibit goldenyellow fluorescence due to the PE. Cells infected with influenza B virus, metapnemovirus or adenovirus 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.
Here's a summary of the acceptance criteria and study details for the D3 FastPoint L-DFA Respiratory Virus Identification Kit, based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance
The document does not explicitly state pre-defined acceptance criteria (e.g., "Sensitivity must be >X%"). Instead, it presents the results of reproducibility and clinical performance studies, implying that these results met the necessary standards for clearance. The "Reported Device Performance" column reflects the results from the clinical sensitivity and specificity studies presented in Tables 5.10-5.15 (for NP wash/aspirate) and 5.16-5.21 (for NP swab). For analytical performance, the reproducibility and LOD results are provided.
| Criterion Type | Acceptance Criteria (Implicit) | Reported Device Performance (Summary) |
|---|---|---|
| Analytical Performance | ||
| Reproducibility | Consistent detection of viral antigens across different sites and runs. | Influenza A/B Reagent: Total agreement 99.3% (278/280). RSV/hMPV Reagent: Total agreement 100% (280/280). HPIV/Adenovirus Reagent: Total agreement 100% (280/280). |
| Limit of Detection (LoD) | Detection of specific viral strains at low concentrations. | Flu A: 50 infected cells/mL Flu B: 50 infected cells/mL RSV: 100 infected cells/mL hMPV A1: 100 infected cells/mL Adenovirus: 100 infected cells/mL HPIV-1: 100 infected cells/mL HPIV-2: 25 infected cells/mL HPIV-3: 50 infected cells/mL |
| Analytical Reactivity (Inclusivity) | Detection of various strains of targeted viruses. | Detected all tested strains of Influenza A (13), Influenza B (7), RSV (3), hMPV (4), HPIV (3), and Adenovirus (10). |
| Clinical Performance (NP Wash/Aspirate) | Acceptable sensitivity and specificity compared to comparator methods. | Flu A: Sensitivity 84.8%, Specificity 99.5% Flu B: Sensitivity 81.8%, Specificity 100.0% RSV: Sensitivity 98.6%, Specificity 99.8% Adenovirus: Sensitivity 92.3%, Specificity 100.0% HPIV: Sensitivity 92.0%, Specificity 99.3% hMPV: Sensitivity 68.8%, Specificity 100.0% |
| Clinical Performance (NP Swab) | Acceptable sensitivity and specificity compared to comparator methods. | Flu A: Sensitivity 87.7%, Specificity 99.8% Flu B: Sensitivity 87.9%, Specificity 99.8% RSV: Sensitivity 97.5%, Specificity 100.0% Adenovirus: Sensitivity 100.0%, Specificity 100.0% (Note: Low prevalence, caution advised) HPIV: Sensitivity 92.9%, Specificity 100.0% hMPV: Sensitivity 54.5%, Specificity 100.0% |
2. Sample Size Used for the Test Set and Data Provenance
-
Clinical Test Set:
- Total Specimens Evaluated: 1519
- Provenance: Prospectively collected excess remnants of respiratory specimens from symptomatic individuals suspected of respiratory infection.
- Country of Origin: 4 geographically diverse U.S. clinical laboratories.
- Retrospective or Prospective: Prospective studies (January 2009 - March 2009).
- Specific Breakdown for Clinical Performance Tables:
- NP Wash/Aspirate (Sites 1, 2, and 3 combined): Number of specimens varies per virus (e.g., 637 for Flu A, 694 for hMPV).
- NP Swab (Sites 3 and 4 combined): Number of specimens varies per virus (e.g., 689 for Flu A, 675 for hMPV).
-
Analytical Test Set (Reproducibility):
- Sample Size: 5 randomized panel members for each of the 3 panels (Influenza A/B, RSV/hMPV, HPIV/Adenovirus). Each panel was tested daily in two separate runs for 5 days by four different laboratories (40 total runs per virus group). This means 40 replicates for each panel member for a given virus.
- Provenance: Proficiency-level antigen control slides with infected cells.
-
Analytical Test Set (Limit of Detection):
- Sample Size: 10 replicate microscope slides for each dilution level of 8 characterized respiratory virus isolates.
- Provenance: Dilution series of infected model cells.
-
Analytical Test Set (Analytical Reactivity/Inclusivity):
- Sample Size: Low concentration infected cell suspensions (approximately 4% cells infected, 25-50 infected cells) for various viral strains for each of the 3 reagents.
- Provenance: Culture isolates of various influenza A (13 strains), influenza B (7 strains), RSV (3 strains), hMPV (4 strains), HPIV (3 strains), and Adenovirus (10 strains).
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts
The document does not specify the number or qualifications of experts used to establish the ground truth for the clinical test set. Instead, it defines "True" positive and "True" negative based on a composite comparator method:
- For Influenza A, Influenza B, RSV, Parainfluenza, and Adenovirus: Direct Specimen Fluorescent Antibody (DSFA) test with an FDA cleared device, and viral culture confirmation of all negatives (as determined by the comparator DSFA test).
- For Human Metapneumovirus (hMPV): 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.
4. Adjudication Method for the Test Set
The adjudication method used for establishing the ground truth for the clinical test set was a composite comparator method. This means results from multiple established methods (DSFA, viral culture, and for hMPV, RT-PCR with sequencing) were combined to determine the "true" status of a specimen. There is no mention of a specific expert panel adjudication method like "2+1" or "3+1" for interpreting these comparator results in case of discrepancies; rather, the definition of "true" positive/negative indicates the hierarchy of methods (e.g., positive by DSFA or viral culture is "true" positive).
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance
No MRMC comparative effectiveness study was done. This device is a diagnostic kit read by a human using a fluorescence microscope, but the study focuses on the kit's performance against comparator methods, not on human reader improvement with or without AI assistance.
6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) was done
This is not applicable as the D3 FastPoint L-DFA Respiratory Virus Identification Kit is a diagnostic kit that relies on a human reading the results under a fluorescence microscope. It is not an AI algorithm. The performance presented is of the kit as used by a human.
7. The Type of Ground Truth Used
The ground truth used for the clinical test set was a composite comparator method, which included:
- FDA cleared Direct Specimen Fluorescent Antibody (DSFA) tests
- Viral Culture
- Validated hMPV real-time RT-PCR followed by bi-directional sequencing analysis (for hMPV only)
- NCBI GenBank database matching with acceptable E-values for bi-directional sequencing data.
8. The Sample Size for the Training Set
The document does not explicitly mention a "training set" in the context of machine learning or AI development. This device is a diagnostic reagent kit, not an AI algorithm. The studies conducted are for analytical and clinical validation of the kit itself.
9. How the Ground Truth for the Training Set was Established
As there is no "training set" in the context of this diagnostic kit, this question is not applicable. The ground truth for the comparator methods was established using established laboratory techniques and FDA-cleared devices, as described in point 7.
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D3 FastPoint L-DFA Respiratory Virus Identification Kit
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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:
April 17, 2009
Device Name:
Trade name - D3 FastPoint L-DFA Respiratory Virus Identification Kit Common name - Respiratory virus DFA assay Classification name - Antisera, Cf, Influenza Virus A, B, C Product Code - GNW Regulation - 21 CFR 866.3330, Class I, Influenza virus serological reagents; 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 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
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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.
D3 Duet DFA RSV/Respiratory Virus Screening Kit (K081928)
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 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
{2}------------------------------------------------
attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
D3 DFA Metapneumovirus Identification Kit (K090073)
The Diagnostic Hybrids, Inc. device, D3 DFA Metapneumovirus Identification Kit (DJ 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 FDAcleared hMPV molecular assay.
Device Description:
The D3 FastPoint L-DFA Respiratory Virus Identification Kit uses three blends (each called a "L-DFA Reagent") of viral antigen-specific murine monoclonal antibodies that are directly labeled with either R-PE (influenza A virus, respiratory syncytial virus, and parainfluenza virus) or fluorescein (influenza B virus, metapneumovirus, and adenovirus) for the rapid identification of respiratory viruses in nasal and nasopharyngeal swabs and aspirates from patients with signs and symptoms of respiratory infection.
Kit Components:
-
- D3 FastPoint L-DFA Influenza A/Influenza B Reagent, 4.0-mL. One dropper bottle containing a mixture of PE-labeled murine monoclonal antibodies directed against influenza A virus antigens and FITC-labeled murine monoclonal antibodies directed against influenza B virus antigens. The buffered, stabilized, aqueous solution contains Evans Blue and propidium iodide as counter-stains and 0.1% sodium azide as preservative.
-
- D3 FastPoint L-DFA RSV/MPV Reagent, 4.0-mL. One dropper bottle containing a mixture of PE-labeled murine monoclonal antibodies directed against respiratory syncytial virus antigens and FITC-labeled murine monoclonal antibodies directed against metapneumovirus antigens. The buffered, stabilized, aqueous solution contains Evans Blue and propidium iodide as counter-stains and 0.1% sodium azide as preservative.
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-
- D3 FastPoint L-DFA PIV/Adenovirus Reagent, 4.0-mL. One dropper bottle containing a mixture of PE-labeled murine monoclonal antibodies directed against parainfluenza virus types 1, 2, or 3 antigens and FITClabeled murine monoclonal antibodies directed against adenovirus 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 4% sodium azide (0.1% sodium azide after dilution to 1X using de-mineralized water).
-
- Re-suspension Buffer, 6.0-mL. One bottle of a buffered glycerol solution and 0.1% sodium azide.
-
- D3 FastPoint L-DFA Respiratory Virus Antigen Control Slides, 5-slides. Five individually packaged control slides containing 6 wells with cell culture-derived positive and negative control cells. Each positive well is identified as to the virus infected cells present, i.e., influenza A virus, influenza B virus, respiratory syncytial virus, metapneumovirus, parainfluenza virus, and adenovirus. The negative wells contain noninfected cells. Each slide is intended to be stained only one time.
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 in 3 separate vials, each containing one of the 3 above reagents. 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 influenza A virus, respiratory syncytial virus, or parainfluenza virus types 1, 2 and 3 will exhibit goldenyellow fluorescence due to the PE. Cells infected with influenza B virus, metapnemovirus or adenovirus 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.
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Intended Use:
The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA Respiratory Virus Identification Kit is intended for the qualitative identification of influenza A virus, influenza B virus, respiratory syncytial virus, human metapneumovirus, adenovirus and to screen for the presence of parainfluenza virus types 1, 2, and 3 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 influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus or parainfluenza viruses 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 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. Since influenza strains display antigenic drift and shift from year to year, performance characteristics may vary. If infection with a novel influenza A virus is suspected, based on clinical and epidemiological screening criteria communicated by public health authorities, collect specimens following appropriate infection control precautions and submit to state or local health departments, for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.2
Technological Characteristics, Compared to Predicate Device:
| Table 5.1: Characteristics of the D³ FastPoint L-DFA Kit are compared to those of the followingDiagnostic Hybrids (DHI) predicate devices | ||||
|---|---|---|---|---|
| Characteristics | D³ FastPoint L-DFA 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-DFARespiratory VirusIdentification Kit isintended for the | The DiagnosticHybrids, Inc. D³Ultra ™ DFA (directfluorescentantibody)Respiratory Virus | The DiagnosticHybrids, Inc. device,D³ Duet DFARSV/RespiratoryVirus Screening Kit,is intended for the | The DiagnosticHybrids, Inc. deviceD³ DFAMetapneumovirusIdentification Kit, isintended for the |
| Characteristics | D³ FastPoint L-DFA Kit Subject Device | D³ Ultra Kit 510(k) #K061101 | D³ Duet RSV Kit 510(k) # K081928 | D³ MPV Kit 510(k) # K090073 |
| qualitativeidentification ofinfluenza Avirus, influenza Bvirus, respiratorysyncytial virus,humanmetapneumovirus,adenovirus and toscreen for thepresence ofparainfluenza virustypes 1, 2, and 3 innasal andnasopharyngealswabs andaspirates/washesspecimens frompatients with signsand symptoms ofrespiratory infectionby direct detectionofimmunofluorescenceusing monoclonalantibodies (MAbs).It is recommendedthat specimensfound to be negativefor influenza Avirus, influenza Bvirus, respiratorysyncytial virus,adenovirus orparainfluenzaviruses afterexamination of thedirect specimenresult be confirmedby cell culture.Specimens found tobe negative forhumanmetapneumovirusafter examination ofthe direct specimenresults should beconfirmed by anFDA cleared human | Screening & 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. | qualitative 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 virusinfection and shouldnot be used as thesole basis fordiagnosis, treatmentor othermanagementdecisions. | qualitative detectionand identification ofhumanmetapneumovirus(hMPV) in nasal andnasopharyngealswabs andaspirates/washes orcell culture. Theassay detects hMPVantigens byimmunofluorescence using a blend ofthree monoclonalantibodies (MAbs),from patients withsigns and symptomsof acute respiratoryinfection. Thisassay detects but isnot intended todifferentiate the fourecognized geneticsub-lineages ofhMPV.Negative results donot preclude hMPVinfection and shouldnot be used as thesole basis fordiagnosis, treatmentor othermanagementdecisions. It isrecommended thatspecimens found tobe negative afterexamination of thedirect specimenresults be confirmedby an FDA-clearedhMPV molecularassay. | |
| Table 5.1: Characteristics of the D³ FastPoint L-DFA Kit are compared to those of the followingDiagnostic Hybrids (DHI) predicate devices | ||||
| Characteristics | D³ FastPoint L-DFAKit Subject Device | D³ Ultra Kit510(k) #K061101 | D³ Duet RSV Kit510(k) # K081928 | D³ MPV Kit510(k) # K090073 |
| metapneumovirusmolecular assay.Negative results donot precluderespiratory virusinfection and shouldnot be used as thesole basis fordiagnosis, treatmentor othermanagementdecisions. | ||||
| Target Viruses | influenza A virus,influenza B virus,respiratory syncytialvirus,metapneumovirus,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 | 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 Reagentscontain 18 MAbs to8 differentrespiratory viruses(influenza A virus,influenza B virus,respiratory syncytialvirus,metapneumovirus,adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 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 the | Direct labeling, | Direct labeling,- using R-Phycoerythrin (R-PE) to label the | Direct labeling, |
| Table 5.1: Characteristics of the D³ FastPoint L-DFA Kit are compared to those of the followingDiagnostic Hybrids (DHI) predicate devices. | ||||
| Characteristics | D³ FastPoint L-DFAKit Subject Device | D³ Ultra Kit510(k) #K061101 | D³ Duet RSV Kit510(k) # K081928 | D³ MPV Kit510(k) # K090073 |
| A virus, RSV andparainfluenza virustypes 1, 2 and 3. | syncytial virus. | |||
| - using fluoresceinisothiocyanate(FITC) to labelinfluenza B virus,metapneumovirusand adenovirusMAbs withfluorescein. | - using fluoresceinisothiocyanate(FITC) to label allMAbs withfluorescein. | - using fluoresceinisothiocyanate(FITC) to label allother MAbs withfluorescein. | - using fluoresceinisothiocyanate(FITC) to label allMAbs withfluorescein. | |
| R-Phycoerythrin-labeledMAbs | influenza A virus,respiratory syncytialvirus, parainfluenzavirus type 1,parainfluenza virustype 2,parainfluenza virustype 3 | None | respiratory syncytialvirus | None |
| Fluorescein-labeled MAbs | influenza B virus,metapneumovirus,adenovirus | influenza A virus,influenza B virus,respiratory syncytialvirus, adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 3 | influenza A virus,influenza B virus,adenovirus,parainfluenza virustype 1, parainfluenzavirus type 2,parainfluenza virustype 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 | Influenza A and B:The fluorescence iscytoplasmic orbright nuclear orboth. Cells appearround.RespiratorySyncytial Virus:The fluorescence iscytoplasmic. Cellsappear round. | Influenza A and B:The fluorescence iscytoplasmic, nuclearor both.Cytoplasmicstaining is oftenpunctate with largeinclusions whilenuclear staining isuniformly bright.Respiratory | Influenza A andB: Thefluorescence iscytoplasmic,nuclear or both.Cytoplasmicstaining is oftenpunctate with largeinclusions whilenuclear staining isuniformly bright.Respiratory | Metapneumovirus:The fluorescence iscytoplasmic andpunctate with smallinclusions in thesyncytia.Negative: Entirecell fluoresce reddue to the EvansBlue counter-stain. |
2 FDA Guidance Document: In Vitro Diagnostic Devices to Detect Influenza A Viruses: Labeling and Regulatory Path; Issued 4/10/2006
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| Table 5.1: Characteristics of the D FastPoint L-DFA Kit are compared to those of the following | ||||||
|---|---|---|---|---|---|---|
| Diagnostic Hybrids (DHI) predicate devicesD3 FastPoint L-DFA | D3 Ultra Kit | D3 Duet RSV Kit | D' MPV Kit | |||
| Characteristics | Kit Subject Device | 510(k) #K061101 | 510(k) # K081928 | 510(k) # K090073 | ||
| cytoplasmic andpunctate. Cellsappear round.Parainfluenza 1, 2,3: The fluorescenceis cytoplasmic. Cellsappear round.Adenovirus: Thefluorescence iscytoplasmic orbright nuclear orboth. Cells appearround.Negative: Cellsfluoresce red due tothe Evans Bluecounter-stain.Nuclei: Cell Nucleifluoresce orange-reddue to thePropidium Iodidecounter-stain. | cytoplasmic andpunctate with smallinclusions in thesyncytia.Parainfluenza 1, 2,3: The fluorescenceis cytoplasmic andpunctate withirregular inclusions.Types 2 and 3 causethe formation ofsyncytia.Adenovirus: Thefluorescence iscytoplasmic andpunctate or brightnuclear or both.Negative: Cellsfluoresce red due tothe Evans Bluecounter-stain. | The fluorescenceis cytoplasmic andpunctate withsmall inclusions inthe syncytia.Parainfluenza 1,2, 3: Thefluorescence iscytoplasmic andpunctate withirregularinclusions. Types2 and 3 cause theformation ofsyncytia.Adenovirus: Thefluorescence iscytoplasmic andpunctate or brightnuclear or both.Negative: Cellsfluoresce red dueto the Evans Bluecounter-stain. | ||||
| Analytical specificity (forinfluenza A virus strains;MAbs are reactive with alllisted strains) | l 3 influenza Astrains: Influenza AMexico/4108/2009(H1N1) from CDC*Influenza ACalifornia/07/2009(H1N1) from CDC*,Aichi (H3N2), Mal(H1N1), Hong Kong(H3N2), Denver(H1N1), PortChalmers (H3N2),Victoria (H3N2),New Jersey (HINI),WS (HIN1), PR(H1N1), Wisconsin(H3N2), WS(HINI),A/NWS/33 (HIN1) | 10 influenza Astrains: Aichi(H3N2), Mal(H1N1), Hong Kong(H3N2), Denver(H1N1), PortChalmers (H3N2),Victoria (H3N2),New Jersey (H1N1),WS (HINI),PR,(HINI),A/NWS/33 (H1N1) | 10 influenza Astrains: Aichi(H3N2), Mal(H1N1), Hong Kong(H3N2), Denver(H1N1), PortChalmers (H3N2),Victoria (H3N2),New Jersey (HINI),WS (HINI), PR(HIN1), A/NWS/33(HINI) | No reaction wasseen to any of thetested influenza Aviruses with theMetapneumovirusDFA Reagent | ||
| Analytical specificity (forInfluenza B virus strains;MAbs are reactive with alllisted strains) | 7 influenza Bstrains: Hong Kong,Maryland, Mass, | 7 influenza Bstrains: Hong Kong,Maryland, Mass, | 7 influenza Bstrains: Hong Kong,Maryland, Mass, | No reaction wasseen to any of thetested influenza B | ||
| Table 5.1: Characteristics of the D³ FastPoint L-DFA Kit are compared to those of the following .Diagnostic Hybrids (DHI) predicate devices | ||||||
| Characteristics | D³ FastPoint L-DFAKit Subject Device | D³ Ultra Kit510(k) #K061101 | D³ Duet RSV Kit510(k) # K081928 | D³ MPV Kit510(k) # K090073 | ||
| GL, Taiwan,B/Lee/40, Russia | GL, Taiwan,B/Lee/40, Russia | GL, Taiwan,B/Lee/40, Russia | viruses with theMetapneumovirusDFA Reagent | |||
| Analyticalspecificity(cross-reactivitystudies; variousstrains ofmicroorganismsand cell lines) | Viruses | 22 | 31 | 32 | 59 | |
| Bacteria | 22 | 18 | 25 | 25 | ||
| Chlamydiaspp. | 1 | 1 | 3 | 3 | ||
| Yeast | 1 | 0 | 1 | 1 | ||
| Protozoan | 01 | 0 | 1 | 1 | ||
| Cell lines | N/A | 17 | 17 | 16 |
Sec05_FastPoint_L-DFA_Final.doc
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- Although the D2 FastPoint L-DFA Influenza A/Influenza B Reagent has been shown to detect the 2009 HIN1 virus in two culture isolates, the performance characteristics of this device with clinical specimens that are positive for the 2009 HINI influenza virus have not been established. The D2 FastPoint L-DFA Influenza A/Influenza B DFA Reagent can distinguish between influenza A and B viruses, but it cannot differentiate influenza subtypes.
Analytical Performance:
Precision/Reproducibility:
Assay precision, intra-assay variability and inter assay variability were assessed with 3 panels of proficiency-level antigen control slides. Each of the 3 reproducibility panels consisted of 5 randomized panel members.
The Influenza A/B panel consisted of the following:
- a. Low level influenza A (Victoria strain) infected cells.
- b. Low level influenza B (Taiwan strain) infected cells.
- Low level influenza A (Victoria strain) infected cells mixed c. with mid level influenza B (Taiwan strain) infected cells.
- d. Low level influenza B (Victoria strain) infected cells mixed with mid level influenza A (Victoria strain) infected cells.
- Mid level non-infected (negative) cells. e.
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.
- Mid level non-infected (negative) cells. e.
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The HPIV/Adenovirus panel consisted of the following:
- a. Low level Para 1 (C-35 strain) infected cells.
- b. Low level Adenovirus (ATCC type 1) infected cells.
- c. Low level Para 1 (C-35 strain) infected cells mixed with mid level Adenovirus (ATCC type 1) infected cells.
- d. Low Adenovirus (ATCC type 1) infected cells mixed with mid level Para 1 (C-35 strain) infected cells.
- Mid level non-infected (negative) cells. e.
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 x108 to 3.5 x108 total cells.
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 D3 FastPoint L-DFAInfluenza A/Influenza B Reagent, the combined data from the four Study Sites demonstrated reproducible detection of influenza A virus by the R-PE labeled MAbs and reproducible detection of influenza B virus by the FITC-labeled MAbs. The presence of influenza A virus infected cells was reported in 100% (120/120) of the wells in which the infected cells were expected. The presence of influenza B virus 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 95% (38/40) of the wells in which infected cells were not present. The total percent agreement for the D3 FastPoint L-DFAa A/Influenza B Reagent was 99.3% (278/280):
| Table 5.2: Reproducibility Study Results using the D³ FastPoint L-DFA Influenza A/Influenza B Reagent | ||||||||
|---|---|---|---|---|---|---|---|---|
| PanelMember | Negative | Flu ALowLevel | Flu BLowLevel | Mixed InfectionFlu A MidLevel | Mixed InfectionFlu B LowLevel | Mixed InfectionFlu A LowLevel | Mixed InfectionFlu B MidLevel | Total %Agreement |
| Concentration | Noinfectedcells | 4 to 10%infectedcells | 4 to 10%infectedcells | 20 to 30%infectedcells | 4 to 10%infectedcells | 4 to 10%infectedcells | 20 to 30%infectedcells | |
| Site 1AgreementwithExpectedresult | 8/10(80%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 68/70(97.1%) |
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D3 FastPoint L-DFA Respiratory Virus Identification Kit
8/24/2009 Section 05, Page 12 of 23
| Site2 | AgreementwithExpectedresult | 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 | AgreementwithExpectedresult | 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 | AgreementwithExpectedresult | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 70/70(100%) |
| TotalAgreementwith Expectedresult | 38/40(95%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 278/280(99.3%) | |
| 95% CI | 83.1 -99.4% | 91.2 -100% | 91.2 -100% | 91.2 -100% | 91.2 -100% | 91.2 -100% | 91.2 -100% | 97.4 -99.9% |
For the D3 FastPoint L-DFA RSV/hMPV 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 D3 FastPoint L-DFA RSV/hMPV Reagent was 100% (280/280):
| Table 5.3: Reproducibility Study Results using the D3 FastPoint L-DFA RSV/hMPV Reagent | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| PanelMember | RSVLowLevel | hMPVLowLevel | Mixed Infection | Mixed Infection | |||||
| Negative | Concentration | Noinfectedcells | 4 to 10%infectedcells | 4 to 10%infectedcells | 20 to 30%infectedcells | 4 to 10%infectedcells | |||
| Site1 | AgreementwithExpectedresult | 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 | AgreementwithExpectedresult | 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 | AgreementwithExpectedresult | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 70/70(100%) |
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Diagnostic Hybrids, Inc.
D' FastPoint L-DFA Respiratory Virus Identification Kit 8/24/2009 Section 05, Page 13 of 23
| Site | AgreementwithExpectedresult | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 70/70(100%) |
|---|---|---|---|---|---|---|---|---|---|
| 4 | TotalAgreementwith Expectedresult | 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% |
For the D3 FastPoint L-DFA HPIV/Adenovirus Reagent, the combined data from the four Study Sites demonstrated reproducible detection of HPIV-1 by the R-PE labeled MAbs and reproducible detection of Adenovirus by the FITC-labeled MAbs. The presence of HPIV-1 infected cells was reported in 100% (120/120) of the wells in which the infected cells were expected. The presence of Adenovirus 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 D3 FastPoint L-DFA HPIV/Adenovirus was 100% (280/280):
| Table 5.4: Reproducibility Study Results using the D³ FastPoint L-DFA HPIV/Adenovirus Reagent | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| PanelMember | Negative | HPIV-1Low Level | AdenovirusLow Level | Mixed Infection | Mixed Infection | Total %Agreement | |||
| Concentration | Noinfectedcells | 4 to 10%infectedcells | 4 to 10%infectedcells | HPIV-1Mid Level | AdenovirusLow Level | HPIV-1Low Level | AdenovirusMid Level | ||
| Site1 | AgreementwithExpectedresult | 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 | AgreementwithExpectedresult | 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 | AgreementwithExpectedresult | 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 | AgreementwithExpectedresult | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 10/10(100%) | 70/70(100%) |
| TotalAgreementwith Expectedresult | 40/40(100%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 40/40(100%) | 280/280(100%) |
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Diagnostic Hybrids, Inc
D3 FastPoint L-DFA Respiratory Virus Identification Kit
8/24/2009 Section 05, Page 14 of 23
| Table 5.4: | Reproducibility Study Results using the D³ FastPoint L-DFA HPIV/Adenovirus Reagent | ||||||
|---|---|---|---|---|---|---|---|
| 95% CI | 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 D3 FastPoint L-DFA Reagents was addressed using dilution series of infected model cells. Model cells for 8 characterized respiratory virus isolates; influenza A virus (ATCC Victoria strain), influenza B virus (ATCC Taiwan strain), respiratory syncytial virus (ATCC Washington strain), adenovirus (ATCC type 1), human metapneumovirus subtype A1 (clinical strain), parainfluenza virus types 1, 2, and 3 (ATCC strains C-35, Greer, and C243 respectively) 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 25uL 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-μL 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 snot 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 5.5 below:
| Table 5.5:Kit | Limit of Detections of the D3 FastPoint L-DFA Respiratory Virus Identification | ||
|---|---|---|---|
| Virus Strain | Infected cells/mL | Number of replicates with positive cells | LOD determination |
| 500 | 10/10 | ||
| 100 | 10/10 | ||
| 50 | 10/10 | ||
| 25 | 5/10 | ||
| Flu A(ATCC Victoria strain) | 12.5 | 3/10 | 50 infected cells/mL |
| 6 | 2/10 | ||
| 3 | 0/10 | ||
| 1.5 | 2/10 | ||
| 0.8 | 0/10 | ||
| 0.4 | 0/10 | ||
| 2000 | 10/10 | ||
| 400 | 10/10 | ||
| 200 | 10/10 | ||
| 100 | 10/10 | ||
| Flu B(ATCC Taiwan strain) | 50 | 10/10 | 50 infected cells/mL |
| 25 | 7/10 | ||
| 12.5 | 4/10 | ||
| 6 | 2/10 | ||
| 3 | 0/10 | ||
| 1.5 | 0/10 | ||
| Table 5.5:Kit | Limit of Detections of the D³ FastPoint L-DFA Respiratory Virus Identification | ||
| Virus Strain | Infected cells/mL | Number of replicates withpositive cells | LOD determination |
| 1000 | 10/10 | ||
| 200 | 10/10 | ||
| 100 | 10/10 | ||
| 50 | 7/10 | ||
| RSV(ATCC Washingtonstrain) | 25 | 7/10 | 100 infected cells/mL |
| 12.5 | 6/10 | ||
| 6 | 1/10 | ||
| 3 | 0/10 | ||
| 1.5 | 0/10 | ||
| 0.8 | 0/10 | ||
| 2000 | 10/10 | ||
| 400 | 10/10 | ||
| 200 | 10/10 | ||
| 100 | 10/10 | ||
| hMPV A1(Clinical strain) | 50 | 6/10 | 100 infected cells/mL |
| 25 | 2/10 | ||
| 12.5 | 0/10 | ||
| 6 | 0/10 | ||
| 3 | 0/10 | ||
| 1.5 | 0/10 | ||
| 1000 | 10/10 | ||
| 200 | 10/10 | ||
| 100 | 9/10 | ||
| 50 | 5/10 | ||
| Adenovirus(ATCC type 1) | 25 | 1/10 | 100 infected cells/mL |
| 12.5 | 0/10 | ||
| 6 | 0/10 | ||
| 3 | 0/10 | ||
| 1.5 | 0/10 | ||
| 0.8 | 0/10 | ||
| 500 | 10/10 | ||
| 100 | 10/10 | ||
| 50 | 6/10 | ||
| 25 | 2/10 | ||
| HPIV-1(ATCC strain C-35) | 12.5 | 1/10 | 100 infected cells/mL |
| 6 | 0/10 | ||
| 3 | 0/10 | ||
| 1.5 | 0/10 | ||
| 0.8 | 0/10 | ||
| 0.4 | 0/10 | ||
| 500 | 10/10 | ||
| 100 | 10/10 | ||
| 50 | 10/10 | ||
| 25 | 9/10 | ||
| HPIV-2(ATCC strain Greer) | 12.5 | 6/10 | |
| (ATCC strain Greer) | 6 | 5/10 | |
| 3 | 3/10 | ||
| 1.5 | 1/10 | ||
| 0.8 | 0/10 | ||
| 0.4 | 0/10 | ||
| HPIV-3(ATCC strain C243) | 1000 | 10/10 | 50 infected cells/mL |
| 200 | 10/10 | ||
| 100 | 10/10 | ||
| 50 | 9/10 | ||
| Table 5.5: Limit of Detections of the D3 FastPoint L-DFA Respiratory Virus IdentificationKit | |||
| Virus Strain | Infected cells/mL | Number of replicates withpositive cells | LOD determination |
| 25 | 6/10 | ||
| 12.5 | 2/10 | ||
| 6 | 0/10 | ||
| 3 | 0/10 | ||
| 1.5 | 0/10 | ||
| 0.8 | 0/10 |
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Sec05_FastPoint_L-DFA_Final.doc
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D3 FastPoint L-DFA Respiratory Virus Identification Kit
8/24/2009 Section 05, Page 16 of 23
Analytical reactivity (inclusivity)
Analytical reactivity (inclusivity) of the D3FastPoint L-DFA Influenza A/Influenza B Reagent was evaluated using 13 influenza A virus and 7 influenza B virus strains. Low concentration infected cell suspensions (approximately 4% cells infected, 25-50 infected cells) were prepared for each viral strain. The suspensions were stained with the D3 FastPoint L-DFA Influenza A/Influenza B Reagent.
| Table 5.6: Analytical Reactivity (inclusivity) of the D³ FastPoint L-DFA Influenza A/Influenza BReagent on various influenza A virus and influenza B virus strains | ||
|---|---|---|
| Influenza Strains | Infected Cell Concentration(as multiples of the respectiveestablished LoD concentration | D³ FastPoint L-DFA Influenza A/Influenza B Reagent Results |
| Influenza A Mexico/4108/2009(H1N1) from CDC* | 20x LoD | 19 Golden-yellow fluorescent cells |
| Influenza A California/07/2009(H1N1) from CDC* | 20x LoD | 26 Golden-yellow fluorescent cells |
| Influenza A Wisconsin/56/2005(H3N2) | 20x LoD | 39 Golden-yellow fluorescent cells |
| Influenza A WS, VR-1520 (H1N1) | 20x LoD | 67 Golden-yellow fluorescent cells |
| Influenza A Hong Kong, VR-544(H3N2) | 20x LoD | 13 Golden-yellow fluorescent cells |
| Influenza A New Jersey, VR-897(H1N1) | 20x LoD | 15 Golden-yellow fluorescent cells |
| Influenza A A/NWS/33(H1N1) | 20x LoD | 10 Golden-yellow fluorescent cells |
| Influenza A Victoria, VR-822 (H3N2) | 20x LoD | 10 Golden-yellow fluorescent cells |
| Influenza A PR, VR-95 (H1N1) | 20x LoD | 20 Golden-yellow fluorescent cells |
| Influenza A Port Chalmers, VR-810(H3N2) | 20x LoD | 8 Golden-yellow fluorescent cells |
| Influenza A Aichi, VR-547 (H3N2) | 20x LoD | 28 Golden-yellow fluorescent cells |
| Influenza A Denver, VR-546 (H1N1) | 20x LoD | 30 Golden-yellow fluorescent cells |
| Influenza A Mal, VR-98 (H1N1) | 20x LoD | 21 Golden-yellow fluorescent cells |
| Influenza B GL/1739/54, VR-103 | 20x LoD | 13 Apple-green fluorescent cells |
| Influenza B Taiwan/2/62, VR-295 | 20x LoD | 44 Apple-green fluorescent cells |
| Influenza B Hong Kong/5/72, VR-823 | 20x LoD | 21 Apple-green fluorescent cells |
| Influenza B Maryland/1/59, VR-296 | 20x LoD | 22 Apple-green fluorescent cells |
| Influenza B Russia, VR-790 | 20x LoD | 36 Apple-green fluorescent cells |
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Diagnostic Hybrids, Inc.
D3 FastPoint L-DFA Respiratory Virus Identification Kit 8/24/2009 Section 05, Page 17 of 23
| Table 5.6: Analytical Reactivity (inclusivity) of the D3, FastPoint L-DFA Influenza A/Influenza BReagent on various influenza A virus and influenza B virus strains | ||
|---|---|---|
| Influenza Strains | Infected Cell Concentration(as multiples of the respectiveestablished LoD concentration | D3 FastPoint L-DFA Influenza A/Influenza B Reagent Results |
| Influenza B B/Lee/40 | 20x LoD | 41 Apple-green fluorescent cells |
| Influenza B Massachusetts, VR-523 | 20x LoD | 67 Apple-green fluorescent cells |
- Although the D3 FastPoint L-DFA Influenza A/Influenza B Reagent has been shown to detect the 2009 H1N1 virus in two culture isolates, the performance characteristics of this device with clinical specimens that are positive for the 2009 H1N1 influenza virus have not been established. The D3 FastPoint L-DFA Influenza A/Influenza B DFA Reagent can distinguish between influenza A and B viruses, but it cannot differentiate influenza subtypes.
Analytical reactivity (inclusivity) of the D3 FastPoint L-DFA RSV/hMPV DFA Reagent was evaluated using 3 RSV virus and 4 hMPV virus strains. Low concentration infected cell suspensions (approximately 4% cells infected, 25-50 infected cells) were prepared for each viral strain. The suspensions were stained with the D FastPoint L-DFA RSV/hMPV Reagent.
| Table 5.7: Analytical Reactivity (inclusivity) of the D3 FastPoint L-DFA RSV/hMPV DFAReagent on various RSV virus and hMPV virus strains | ||
|---|---|---|
| RSV and hMPV Strains | Infected Cell Concentration (asmultiples of the respectiveestablished LoD concentration | D3 FastPoint L-DFA RSV/hMPV ReagentResults |
| 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 |
Analytical reactivity (inclusivity) of the D3 FastPoint L-DFA HPIV/Adenovirus DFA Reagent was evaluated using 3 HPIV virus and 10 Adenovirus strains. Low concentration infected cell suspensions (approximately 4% cells infected, 25-50 infected cells) were prepared for each viral strain. The suspensions were stained with the D FastPoint L-DFA HPIV/Adenovirus Reagent.
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| Table 5.8: | Analytical Reactivity (inclusivity) of the D³ FastPoint L-DFA HPIV/AdenovirusReagent on various HPIV virus and adenovirus strains | |
|---|---|---|
| Parainfluenza and Adenovirus Strains | Infected Cell Concentration(as multiples of the respectiveestablished LoDconcentration | D³ FastPoint L-DFA HPIV/AdenovirusReagent Results |
| Parainfluenza 1 C-35 | 10x LoD | 9 Golden-yellow fluorescent cells |
| Parainfluenza 2 Greer | 10x LoD | 11 Golden-yellow fluorescent cells |
| Parainfluenza 3 C-243 | 10x LoD | 22 Golden-yellow fluorescent cells |
| Adenovirus 1 VR-1 | 10x LoD | 26 Apple-green fluorescent cells |
| Adenovirus 3 VR-3 | 10x LoD | 17 Apple-green fluorescent cells |
| Adenovirus 5 VR-5 | 10x LoD | 15 Apple-green fluorescent cells |
| Adenovirus 6 VR-6 | 10x LoD | 22 Apple-green fluorescent cells |
| Adenovirus 7 VR-7 | 10x LoD | 16 Apple-green fluorescent cells |
| Adenovirus 8 VR-1366 | 10x LoD | 29 Apple-green fluorescent cells |
| Adenovirus 10 VR-1087 | 10x LoD | 34 Apple-green fluorescent cells |
| Adenovirus VR-14 | 10x LoD | 37 Apple-green fluorescent cells |
| Adenovirus Dewitt ATCC Strain | 10x LoD | 15 Apple-green fluorescent cells |
| Adenovirus 31 VR-1109 | 10x LoD | 42 Apple-green fluorescent cells |
Clinical Performance:
Performance of the D3 FastPoint L-DFA Respiratory Virus Identification 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 L-DFA Respiratory Virus Identification Kit was assessed and compared to a predetermined algorithm that used composite comparator methods. The composite comparator methods for influenza A virus, influenza B virus, respiratory syncytial virus, parainfluenza virus, and adenovirus 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
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DSFA test) using a validated3 hMPV real-time RT-PCR followed by bidirectional sequencing analysis comparator assay. The hMPV real-time RT-PCR comparator assay targets the hMPV Nucleocapsid gene. "True" 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-values4 "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 the respiratory viruses within this population as determined by the D3 FastPoint L-DFA Respiratory Virus Identification Kit direct specimen testing is noted in Table 5.9 below:
| Table 5.9: Respiratory Virus Prevalence | |||||||
|---|---|---|---|---|---|---|---|
| Age | TotalSpecimensEvaluated | Flu A# positive(prevalence) | Flu B# positive(prevalence) | RSV# positive(prevalence) | hMPV# positive(prevalence) | Adenovirus# positive(prevalence) | HPIV# positive(prevalence) |
| 0 - 1month | 55 | 0 | 0 | 15 (27.3%) | 2 (3.6%) | 0 | 1 (1.8%) |
| > 1month to2 years | 577 | 27 (4.7%) | 20 (3.5%) | 154 (26.7%) | 41 (7.1%) | 11 (1.9%) | 29 (5.0%) |
| > 2years to12 years | 391 | 43 (11.0%) | 104 (26.6%) | 25 (6.4%) | 17 (4.3%) | 1 (0.3%) | 6 (1.5%) |
| > 12years to21 years | 173 | 19 (11.0%) | 41 (23.7%) | 4 (2.3%) | 3 (1.7%) | 0 | 2 (1.2%) |
| 22 yearsto 30years | 57 | 3 (5.3%) | 14 (24.6%) | 0 | 1 (1.8%) | 0 | 1 (1.8%) |
| 31 yearsto 40years | 71 | 9 (12.7%) | 9 (12.7%) | 1 (1.4%) | 3 (4.2%) | 0 | 0 |
| 41 yearsto 50years | 52 | 5 (9.6%) | 5 (9.6%) | 0 | 1 (1.9%) | 0 | 0 |
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 (TCID30mL) stocks of the 4 hMPV (subtypes A1, A2, B1 and B2) strains diluted in hMPV negative nasopharyngeal clinical matrix, and ranged from 10 - 50 TCIDs/mL.
4 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.
(http://www.ncbi.nlm.nih.gov/books/bv.fcgi?rid=handbook.section.614).
Sec05 FastPoint L-DFA Final.doc
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| Table 5.9: | Respiratory Virus Prevalence | ||||||
|---|---|---|---|---|---|---|---|
| Age | TotalSpecimensEvaluated | Flu A | Flu B | RSV | hMPV | Adenovirus | HPIV |
| # positive(prevalence) | # positive(prevalence) | # positive(prevalence) | # positive(prevalence) | # positive(prevalence) | # positive(prevalence) | ||
| 51 yearsto 60years | 46 | 3 (6.5%) | 3 (6.5%) | 1 (2.2%) | 3 (6.5%) | 0 | 0 |
| 61 yearsto 70years | 33 | 2 (6.1%) | 2 (6.1%) | 1 (3.0%) | 1 (3.0%) | 0 | 0 |
| 71 yearsto 80years | 16 | 2 (12.5%) | 1 (6.3%) | 1 (6.3%) | 4 (25.0%) | 0 | 0 |
| 81 yearsandabove | 7 | 0 | 0 | 1 (14.3%) | 0 | 0 | 1 (14.3%) |
| Age NotReported | 41 | 2 (4.9%) | 14 (34.1%) | 0 | 1 (2.4%) | 1 (2.4%) | 0 |
| Total | 1519 | 115 (7.6%) | 213 (14.0%) | 203 (13.4%) | 77 (5.1%) | 13 (0.9%) | 40 (2.6%) |
- There were seven (7) co-infections detected: 2 - respiratory syncytial virus + metapneumovirus, 2- adenovirus + respiratory syncytial virus, 2- influenza A + metapneumovirus, 1-respiratory syncytial virus + adenovirus and 1-respiratory syncytial virus + parainfluenza virus
Tables 5.10 through 5.15 below show the study results of the NP wash/aspirate specimen type (Sites 1, 2, and 3 combined):
| Table 5-10: Flu A | |||
|---|---|---|---|
| Fresh nasal/nasopharyngeal wash/aspirate | Comparator DSFA (negatives followed by culture with DFA) | ||
| DHI DSFA | Positive | Negative | Total |
| Positive | 56 | 3 | 59 |
| Negative | 10 | 568 | 578 |
| Total | 66 | 571 | 637 |
| 95% CI | |||
| Sensitivity | 56/66 | 84.8% | 73.9-92.5% |
| Specificity | 568/571 | 99.5% | 98.5-99.9% |
| Table 5.1: Flu B | |
|---|---|
| Fresh nasal/nasopharyngeal | Of limited use, difficult to interpret, and low sensitivity |
| wash/aspirate | Comparator DSFA (negatives followed by culture with DFA) | |||
|---|---|---|---|---|
| DHI DSFA | Positive | Negative | Total | |
| Positive | 9 | 0 | 9 | |
| Negative | 2 | 617 | 619 | |
| Total | 11 | 617 | 628 | |
| 95% CI | ||||
| Sensitivity | 9/11 | 81.8% | 48.2-97.7% | |
| Specificity | 617/617 | 100.0% | 99.4-100% |
Sec05_FastPoint_L-DFA_Final.doc
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D³ FastPoint L-DFA Respiratory Virus Identification Kit
Section 05, Page 21 of 23
| Fresh nasal/nasopharyngealwash/aspirate | Comparator DSFA (negatives followed by culture withDFA) | ||
|---|---|---|---|
| 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% |
| ST . BRPT. 1221111 74611:11. 1. 11244211. | Cable State Cable S. 13:40 Adenovirus Adenovirus | .. 19 . 15 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 . 19 |
|---|---|---|
| 1000 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 1STATE CHARE START CO |
| Fresh nasal/nasopharyngealwash/aspirate | Comparator DSFA (negatives followed by culture withDFA) | ||
|---|---|---|---|
| DHI DSFA | Positive | Negative | Total |
| Positive | 12 | 0 | 12 |
| Negative | 1 | 619 | 620 |
| Total | 13 | 619 | 632 |
| 95% CI | |||
| Sensitivity | 12/13 | 92.3% | 64.0-99.8% |
| Specificity | 619/619 | 100.0% | 99.4-100% |
| Table 5.14: HPIV | |||
|---|---|---|---|
| Fresh nasal/nasopharyngealwash/aspirate | Comparator DSFA (negatives followed by culture withDFA) | ||
| DHI DSFA | Positive | Negative | Total |
| Positive | 23 | 4 | 27 |
| Negative | 2 | 599 | 601 |
| Total | 25 | 603 | 628 |
| 95% CI | |||
| Sensitivity | 23/25 | 92.0% | 74.0-99.0% |
| Specificity | 599/603 | 99.3% | 98.3-99.8% |
Table 5.15: Table 5.15: Table 5.15: hMPV Sale Sale Sale ﺍﻟﻤﺴﺘﻘﻠﺔ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘ ﺍﻟﻤﻮﺍﻗﻊ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘ
| Fresh nasal/nasopharyngealwash/aspirate | Comparator DSFA (negatives confirmed by a validatedhMPV real-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 |
| Sensitivity | 55/80 | 68.8% | 57.4-78.7% |
| Specificity | 614/614 | 100.0% | 99.4-100% |
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Tables 5.16 through 5.21 below show the study results of the NP swab specimen type (Sites 3 and 4 combined):
| Table 5.16: Flu A | ||||
|---|---|---|---|---|
| Fresh nasal/nasopharyngeal swab | Comparator DSFA (negatives followed by culture withDFA) | |||
| DHI DSFA | Positive | Negative | Total | |
| Positive | 57 | 1 | 58 | |
| Negative | 8 | 623 | 631 | |
| Total | 65 | 624 | 689 | |
| 95% CI | ||||
| Sensitivity | 57/65 | 87.7% | 77.2-94.5% | |
| Specificity | 623/624 | 99.8% | 99.1-100% |
| Table 5.17: Flu B. | |||
|---|---|---|---|
| Fresh nasal/nasopharyngeal swab | Comparator DSFA (negatives followed by culture with DFA) | ||
| DHI DSFA | Positive | Negative | Total |
| Positive | 203 | 1 | 204 |
| Negative | 28 | 478 | 506 |
| Total | 231 | 479 | 710 |
| 95% CI | |||
| Sensitivity | 203/231 | 87.9% | 83.7-92.1% |
| Specificity | 478/479 | 99.8% | 98.8-100% |
| Table 5.18: RSV | |||
|---|---|---|---|
| Fresh nasal/nasopharyngeal swab | Comparator DSFA (negatives followed by culture with DFA) | ||
| DHI DSFA | Positive | Negative | Total |
| Positive | 39 | 0 | 39 |
| Negative | 1 | 646 | 647 |
| Total | 40 | 646 | 686 |
| 95% CI | |||
| Sensitivity | 39/40 | 97.5% | 86.8-99.9% |
| Specificity | 646/646 | 100.0% | 99.4-100% |
| Table 5.19: | Adenovirus | ||
|---|---|---|---|
| Fresh nasal/nasopharyngeal swab | Comparator DSFA (negatives followed by culture withDFA) | ||
| DHI DSFA | Positive | Negative | Total |
| Positive | 1 | 0 | 1 |
| Negative | 0 | 679 | 679 |
| Total | 1 | 679 | 680 |
Sec05_FastPoint_L-DFA_Final.doc
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8/24/2009 Section 05, Page 23 of 23
| 95% CI | |||
|---|---|---|---|
| Sensitivity | 1/1 | 100.0% | NA |
| Specificity | 679/679 | 100.0% | 99.5-100% |
Note: The sensitivity performance of the D FastPoint L-DFA Respiratory Virus ID Kit detecting adenovirus from direct nasal/nasopharyngeal swab specimens has not been adequately established in the clinical study due to low adenovirus prevalence at the clinical study sites. However, the same MAb pool for adenovirus was validated in previous clinical trials for a number of FDA cleared DSFA devices. Users may wish to further evaluate the sensitivity performance of this kit detecting adenovirus using prospective nasal/nasopharyngeal swab samples.
| Table 5.20: | HPIV | ||
|---|---|---|---|
| Fresh nasal/nasopharyngeal swab | Comparator DSFA (negatives followed by culture withDFA) | ||
| DHI DSFA | Positive | Negative | Total |
| Positive | 13 | 0 | 13 |
| Negative | 1 | 667 | 668 |
| Total | 14 | 667 | 681 |
| 95% CI | |||
| Sensitivity | 13/14 | 92.9% | 66.1-99.8% |
| Specificity | 667/667 | 100.0% | 99.4-100% |
| Table 5.21: hMPV | |||
|---|---|---|---|
| Fresh nasal/nasopharyngeal swab | Comparator DSFA (negatives confirmed by a validatedhMPV real-time RT-PCR followed by bi-directionalsequencing analysis comparator assay) | ||
| DHI DSFA | Positive | Negative | Total |
| Positive | 24 | 0 | 24 |
| Negative | 20 | 631 | 651 |
| Total | 44 | 631 | 675 |
| 95% CI | |||
| Sensitivity | 24/44 | 54.5% | 38.8-69.9% |
| Specificity | 631/631 | 100.0% | 99.4-100% |
Overall at the four Study Sites, the performance results of the D3 FastPoint L-DFA Respiratory Virus Identification Kit, when compared to those of the comparator devices, D3 Ultra Kit, D3 Duet RSV Kit, and D3 Metapneumovirus DFA Reagent, demonstrate that the devices detect respiratory virus antigens in a similar manner.
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Image /page/23/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES. USA" around the perimeter. Inside the circle is a stylized eagle-like symbol with three curved lines representing its wings or feathers.
DEPARTMENT OF HEALTH & HUMAN SERVICES
Public Health Service
Food and Drug Administration 10903 New Hampshire Avenue Building 66 Silver Spring, MD 20993
Mr. Ronald Lollar
SEP 112009
Senior Director, Product Realization, Management, and Marketing Diagnostic Hybrids Inc. 1055 East State Street Suite 100 Athens, OH 45701
Re: K091171
Trade/Device Name: D3 FastPoint L-DFA Respiratory Virus Identification Kit Regulation Number: 21 CFR 866.3980
Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: Class II
Product Code: OMG, LKT, GNX, GOS, GNY
Dated: September 2, 2009
Received: September 8, 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
{24}------------------------------------------------
limited to: registration and listing (21 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) 594-3084. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). Other general information on your responsibilities under the Act may be obtained 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/dsma/dsmamain.html.
Sincerely yours.
Ube Scrf. fir
Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known): K091171
Device Name: D FastPoint L-DFA Respiratory Virus Identification Kit
Indications For Use:
The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA Respiratory Virus Identification Kit is intended for the qualitative identification of influenza A virus, influenza B virus, respiratory syncytial virus, human metapneumovirus, adenovirus and to screen for the presence of parainfluenza virus types 1, 2, and 3 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 influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus or parainfluenza viruses 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 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. Since influenza strains display antigenic drift and shift from year to year, performance characteristics may vary. If infection with a novel influenza A virus is suspected, based on clinical and epidemiological screening criteria communicated by public health authorities, collect specimens following appropriate infection control precautions and submit to state or local health departments, for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.2
Prescription Use X (Part 21 CFR 801 Subpart D) AND/OR
Over-The-Counter Use (21 CFR 807 Subpart C)
ﺎ ﺍﻟﻤﺮﺍﺟﻊ
Division Sign-Off
Office of In Vitro Diagnostic Device Page 1 of Evaluation and Safety
O(K) ki 1171
FDA Guidance Document: In Vitro Diagnostic Devices to Detect Influenza A Viruses: Labeling and Regulatory Path; Issued 4/10/2006
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Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)
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§ 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.