(187 days)
The Diagnostic Hybrids, Inc. device, D3 Duet DFA Influenza A/Respiratory Virus Screening Kit, is intended for the qualitative detection and identification of influenza A, while screening for influenza B virus, respiratory syncytial 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. 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 attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
The Diagnostic Hybrids, Inc. device, D Duet DFA Influenza A/Respiratory Virus Screening Kit, uses a blend of viral antigen-specific murine MAbs. MAbs for influenza A virus are directly labeled with R-phycoerythrin (R-PE) for the rapid detection and identification of influenza A virus. MAbs for influenza B virus, respiratory syncytial virus, adenovirus, and parainfluenza virus types 1, 2, and 3 are directly labeled with fluorescein isothiocyanate (FITC), for rapid detection of these agents.
Kit components:
- D3 Duet DFA Influenza A/Respiratory Virus Screening Reagent Rphycoerythrin-labeled murine MAbs directed against influenza A virus and a mixture of fluorescein-labeled murine MAbs directed against influenza B. respiratory syncytial virus, adenovirus, and parainfluenza virus types 1, 2, and 3. The buffered, stabilized, aqueous solution also contains Evans Blue as a counterstain and 0.1% sodium azide as preservative.
- Normal Mouse Gamma Globulin DFA Reagcnt a mixture of fluorescein labeled murine gamma globulin that has been shown to be non-reactive with any of the listed respiratory viruses. The buffered, stabilized, aqueous solution contains Evans Blue as a counter-stain and 0.1% sodium azide as preservative.
- Respiratory Virus Antigen Control Slides five individually packaged control slides containing wells with cell culture-derived positive and negative control cells. Each positive well is identified with the virus infected cells present, i.e., influenza A virus, influenza B virus, respiratory syncvtial virus, adenovirus, and parainfluenza virus types 1, 2 and 3. The negative well contains uninfected cultured cells. Each slide is intended to be stained only one time.
- Wash Solution Concentrate a 40X concentrate consisting of Tween 20 and 4% sodium azide (0.1% sodium azide after dilution in de-mineralized water) in a 40X phosphate buffered saline solution.
- Mounting Fluid an aqueous, buffered, stabilized solution of glycerol and . 0.1% sodium azide.
The cells to be tested, derived from a clinical specimen or cell culture, are placed onto a glass slide and allowed to air dry. The cells are fixed in acetone. The D3 Duet DFA Influenza A/Respiratory Virus Screening Reagent is added to the cells which are then incubated for 15 to 30 minutes at 35° to 37°C in a humidified chamber or humidified incubator. The stained cells are then washed with the diluted wash solution, a drop of the supplied Mounting Fluid is added and a coverslip is placed on the prepared cells. The cells are examined using a fluorescence microscope. The influenza A virus infected cells will fluoresce golden-yellow, while cells infected with any of the other six viruses will fluoresce apple-green. Uninfected cells will contain no fluorescence but will be stained rcd by the Evans Blue counter-stain. If only golden-yellow fluorescent cells are present the specimen can be reported as positive for influenza A antigen. If only apple-green fluorescent cells are present, the particular virus may be identified using the individual reagents from the DS Ultra™ DFA Respiratory Virus Screening & ID Kit (D3 Ultra) on new, separate cell preparations. If both golden-yellow and apple-green are present, the additional virus may be identified using the individual reagents from the D3 Ultra on new, separate cell preparations.
Here's a breakdown of the acceptance criteria and the study details for the D3 Duet DFA Influenza A/Respiratory Virus Screening Kit, based on the provided text:
Acceptance Criteria and Device Performance
The acceptance criteria for the D3 Duet DFA Influenza A/Respiratory Virus Screening Kit are established by demonstrating substantial equivalence to a legally marketed predicate device, the D3 Ultra DFA Respiratory Virus Screening & ID Kit (K061101). The performance is measured primarily through Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) compared to the predicate device.
Table 1: Acceptance Criteria and Reported Device Performance (Clinical Study - Direct Specimens)
| Target Virus(es) | Criterion (compared to D3 Ultra Final Identification) | Reported Device Performance (D3 Duet with 95% CI) |
|---|---|---|
| Influenza A Virus | Positive Percent Agreement (PPA) >= 95% | 99% (94.5%, 99.8%) |
| Negative Percent Agreement (NPA) >= 99% | 100% (99.7%, 100%) | |
| Influenza B, RSV, Adeno, Para 1, 2, 3 | Positive Percent Agreement (PPA) >= 99% | 100% (99.0%, 100%) |
| Negative Percent Agreement (NPA) >= 99% | 100% (99.5%, 100%) | |
| Individual Virus Performance | Sensitivity >= 90% (where applicable) | Influenza B: 100% (74.12, 100) Adenovirus: 100% (93.1, 100) Parainfluenza type 1: 100% (51.0, 100) Parainfluenza type 2: 100% (20.1, 100) Parainfluenza type 3: 100% (83.2, 100) Respiratory Syncytial Virus: 100% (98.7, 100) |
| Specificity >= 99% (where applicable) | Influenza B, Adeno, Para 1, 2, 3, RSV: All 100% (99.6% to 99.7%, 100%) |
Table 2: Acceptance Criteria and Reported Device Performance (Clinical Study - Cultured Specimens)
| Target Virus(es) | Criterion (compared to D3 Ultra Final Identification) | Reported Device Performance (D3 Duet with 95% CI) |
|---|---|---|
| Influenza A Virus | Positive Percent Agreement (PPA) >= 95% | 100% (94.6%, 100%) |
| Negative Percent Agreement (NPA) >= 99% | 100% (98.4%, 100%) | |
| Influenza B, RSV, Adeno, Para 1, 2, 3 | Positive Percent Agreement (PPA) >= 99% | 100% (95.0%, 100%) |
| Negative Percent Agreement (NPA) >= 99% | 100% (98.4%, 100%) |
Study Details
2. Sample Size Used for the Test Set and Data Provenance
- Direct Fresh Clinical Specimens:
- Sample Size: 1184 specimens (from an initial 1203, with 19 excluded).
- Data Provenance: Prospective study conducted at three unnamed clinical sites. The country of origin is implicitly the United States as it is reviewed by the FDA.
- Cultured Clinical Specimens:
- Sample Size: 298 frozen specimens.
- Data Provenance: Conducted at a fourth unnamed clinical site (Study Site 4) using frozen specimens that were processed for cell culture. The specimens were "derived from nasopharyngeal specimens." The country of origin is implicitly the United States.
3. Number of Experts Used to Establish Ground Truth for the Test Set and Qualifications
The ground truth for the clinical studies (both direct and cultured specimens) was established by comparison to a "cleared DSFA device" (D3 Ultra DFA Respiratory Virus Screening & ID Kit). While clinical experts read the comparator and subject device results, the document does not specify the number of experts used for establishing the ground truth or their qualifications beyond implying standard laboratory personnel performing the D3 Ultra assay. The D3 Ultra itself would have been interpreted by trained laboratory professionals.
4. Adjudication Method for the Test Set
The document describes comparing the D3 Duet results to the "D3 Ultra Final Identification." This implies that the D3 Ultra's result was considered the reference standard or adjudicated ground truth for the clinical studies. There is no mention of a separate adjudication panel or consensus process for discrepancies between the D3 Duet and D3 Ultra results; the D3 Ultra's result is taken as the benchmark.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
No MRMC comparative effectiveness study was done to assess the effect size of human readers improving with AI vs. without AI assistance. The study compares the performance of the new device (D3 Duet) to a predicate device (D3 Ultra) in a standalone manner, with both being manual assays performed by laboratory personnel.
6. Standalone (Algorithm Only) Performance
Yes, the studies evaluate the D3 Duet as a standalone diagnostic device. It is a manual immunofluorescence assay performed by laboratory technicians, not an AI algorithm. Its performance is measured directly against the predicate device.
7. Type of Ground Truth Used
- For Clinical Studies (Direct and Cultured Specimens): The ground truth was the "D3 Ultra Final Identification" results. The D3 Ultra is also an immunofluorescence assay.
- For Analytical Studies (Sensitivity, Specificity, Reactivity): The ground truth was established by prepared viral stocks with known concentrations (TCID50) and identification of specific virus strains.
8. Sample Size for the Training Set
The document does not explicitly mention a "training set" in the context of machine learning or AI. This device is a manual laboratory test using monoclonal antibodies, not an AI system. The analytical and precision/reproducibility studies can be considered as validation work that informs the device's design and reliability before full clinical evaluation.
9. How the Ground Truth for the Training Set was Established
As this is not an AI/ML device, there isn't a "training set" in that sense. The development of the D3 Duet's monoclonal antibodies and reagents would have involved extensive laboratory testing and characterization against known viral isolates and cultures to establish their specificity and reactivity, analogous to establishing ground truth for individual components during development. These methods are detailed in the analytical specificity and reactivity sections, where viral strains and dilutions are tested.
{0}------------------------------------------------
D3 DUET DFA INFLUENZA A/RESPIRATORY VIRUS SCREENING KIT
SECTION 05, 510(K) SUMMARY
Applicant:
DIAGNOSTIC HYBRIDS, INC. 1055 East State Street Suite 100 Athens, OHIO 45701
DEC 2 3 2008
Contact Information:
Gail R. Goodrum Vice President, Regulatory Affairs E-mail: goodrum(@dhiusa.com Telephone: 740-589-3300 Desk Extension: 740-589-3380 FAX: 740-593-8437
Date of preparation of 510(k) summary:
June 13, 2008
Device Name:
Trade name - D3 Duet DFA Influenza A/Respiratory Virus Screening Kit Common name - Fluorescent antibody test for screening Influenza A 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 device to which equivalence is claimed:
K061101, D3 Ultra DFA Respiratory Virus Screening & ID Kit
Intended Use: The Diagnostic Hybrids, Inc. device, D3 Duet DFA Influenza A/Respiratory Virus Screening Kit, is intended for the qualitative detection and identification of influenza A, while screening for influenza B virus, respiratory syncytial 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.
{1}------------------------------------------------
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. 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 attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
Device Description:
The Diagnostic Hybrids, Inc. device, D Duet DFA Influenza A/Respiratory Virus Screening Kit, uses a blend of viral antigen-specific murine MAbs. MAbs for influenza A virus are directly labeled with R-phycoerythrin (R-PE) for the rapid detection and identification of influenza A virus. MAbs for influenza B virus, respiratory syncytial virus, adenovirus, and parainfluenza virus types 1, 2, and 3 are directly labeled with fluorescein isothiocyanate (FITC), for rapid detection of these agents.
Kit components:
- · D3 Duet DFA Influenza A/Respiratory Virus Screening Reagent Rphycoerythrin-labeled murine MAbs directed against influenza A virus and a mixture of fluorescein-labeled murine MAbs directed against influenza B. respiratory syncytial virus, adenovirus, and parainfluenza virus types 1, 2, and 3. The buffered, stabilized, aqueous solution also contains Evans Blue as a counterstain and 0.1% sodium azide as preservative.
- Normal Mouse Gamma Globulin DFA Reagcnt a mixture of fluorescein labeled murine gamma globulin that has been shown to be non-reactive with any of the listed respiratory viruses. The buffered, stabilized, aqueous solution contains Evans Blue as a counter-stain and 0.1% sodium azide as preservative.
- · Respiratory Virus Antigen Control Slides five individually packaged control slides containing wells with cell culture-derived positive and negative control cells. Each positive well is identified with the virus infected cells present, i.e., influenza A virus, influenza B virus, respiratory syncvtial virus, adenovirus, and parainfluenza virus types 1, 2 and 3. The negative well contains uninfected cultured cells. Each slide is intended to be stained only one time.
- · Wash Solution Concentrate a 40X concentrate consisting of Tween 20 and 4% sodium azide (0.1% sodium azide after dilution in de-mineralized water) in a 40X phosphate buffered saline solution.
{2}------------------------------------------------
- Mounting Fluid an aqueous, buffered, stabilized solution of glycerol and . 0.1% sodium azide.
The cells to be tested, derived from a clinical specimen or cell culture, are placed onto a glass slide and allowed to air dry. The cells are fixed in acetone. The D3 Duet DFA Influenza A/Respiratory Virus Screening Reagent is added to the cells which are then incubated for 15 to 30 minutes at 35° to 37°C in a humidified chamber or humidified incubator. The stained cells are then washed with the diluted wash solution, a drop of the supplied Mounting Fluid is added and a coverslip is placed on the prepared cells. The cells are examined using a fluorescence microscope. The influenza A virus infected cells will fluoresce golden-yellow, while cells infected with any of the other six viruses will fluoresce apple-green. Uninfected cells will contain no fluorescence but will be stained rcd by the Evans Blue counter-stain. If only golden-yellow fluorescent cells are present the specimen can be reported as positive for influenza A antigen. If only apple-green fluorescent cells are present, the particular virus may be identified using the individual reagents from the DS Ultra™ DFA Respiratory Virus Screening & ID Kit (D3 Ultra) on new, separate cell preparations. If both golden-yellow and apple-green are present, the additional virus may be identified using the individual reagents from the D3 Ultra on new, separate cell preparations.
Technological Characteristics:
The DHI device, D3 Duet, has been compared directly to the DHI device, D3 Ultra, as the legally marketed device. The technology used in both devices is based on a standard immunofluorescence assay technique utilizing either R-PF. or FITC-labeled MAbs. A summary is provided in Table 5.1 below:
| TABLE 5.1: Technological Characteristics Comparison | |||
|---|---|---|---|
| Characteristic | D3 Duet DFA Influenza A/Respiratory Virus Screening Kit | D3 Ultra DFA Respiratory VirusScreening & ID Kit | |
| Monoclonal antibodies (MAbs) | The Influenza A/Respiratory VirusDFA Screening Reagentcontains 12 MAbs to 6 differentrespiratory viruses (influenza B virus,respiratory syncytial virus, adenovirus,parainfluenza virus type 1,parainfluenza virus type 2,parainfluenza virus type 3), plus 2MAbs to influenza A virus.One of the 2 MAbs to influenza Avirus is different from either of thoseused in the D3 Ultra Reagent; thesecond is the same. | The Respiratory Virus DFA ScreeningReagentcontains 12 MAbs to 6 differentrespiratory viruses (influenza B virus,respiratory syncytial virus, adenovirus,parainfluenza virus type 1,parainfluenza virus type 2,parainfluenza virus type 3), plus 2MAbs to influenza A virus. | |
| TABLE 5.1: Technological Characteristics Comparison | |||
| Characteristic | D3 Duet DFA Influenza A/Respiratory Virus Screening Kit | D3 Ultra DFA Respiratory VirusScreening & ID Kit | |
| Labeling method | Direct labeling,- using R-phycoerythrin (R-PE) tolabel the MAbs to influenza A virusantigens- using fluorescein isothiocyanate(FITC) to label all other MAbs withfluorescein moiety | Direct labeling,- using fluorescein isothiocyanate(FITC) to label all other MAbs withfluorescein moiety | |
| Fluorescein-labeled MAbs | Influenza B virus, respiratory syncytialvirus, adenovirus, parainfluenza virustype 1, parainfluenza virus type 2,parainfluenza virus type 3 | Influenza A virus, influenza B virus,respiratory syncytial virus, adenovirusparainfluenza virus type 1,parainfluenza virus type 2,parainfluenza virus type 3 | |
| Phycoerythrin-labeled MAbs | Influenza A virus(Phycoerythrin-labeled influenza Avirus MAbs stain with golden-yellowfluorescence) | None(Fluorescein-labeled influenza A virusMAbs stain with apple-greenfluorescence) | |
| Cell Fixative | Cell Fixative is the same for both devices:Acetone | ||
| Performance characteristics | |||
| Staining patterns | Staining patterns are the same for both devices:Influenza A and B: The fluorescence is cytoplasmic, nuclear or both.Cytoplasmic staining is often punctate with large inclusions while nuclearstaining is uniformly bright.Respiratory Syncytial Virus: The fluorescence is cytoplasmic and punctatewith small inclusions in the syncytia.Parainfluenza 1, 2, 3: The fluorescence is cytoplasmic and punctate withirregular inclusions. Types 2 and 3 cause the formation of syncytia.Adenovirus: The fluorescence is cytoplasmic and punctate or bright nuclear orboth. | ||
| Analytical sensitivity, accordingto 96-well cell culture platesinfected with Flu A diluted togive a TCID50 of 1 per 0.2-mLinoculum (reported as averageof 4 runs) | There is no significant difference between the two devices for analyticalsensitivity.34.3 ± 12.0culture positives out of 96 | ||
| 34.8 + 9.7culture positives out of 96 | |||
| Analytical specificity (forinfluenza A virus strains; MAbsare reactive with all listedstrains) | Mabs to influenza A virus were shown to be reactive with these virus strains: | ||
| 9 Flu A strains (Aichi, VR-547(H3N2); Mal, VR-98 (H1N1); HongKong, VR-544 (I13N2); Denver, VR-546 (H1N1); Port Chalmers, VR-810(H3N2); Victoria, VR-822 (H3N2);New Jersey, VR-897(H1N1); WS, VR-1520 (H1N1); PR, VR-95 (H1N1)) | 9 Flu A strains (Aichi, VR-547(H3N2); Mal, VR-98 (H1N1); HongKong, VR-544 (H3N2); Denver, VR-546 (H1N1); Port Chalmers, VR-810(H3N2); Victoria, VR-822 (H3N2);New Jersey, VR-897(H1N1); WS, VR-1520 (H1N1); PR, VR-95 (H1N1)) | ||
| Analyticalspecificity (crossreactivity studies;various strains ofmicroorganisms | Device Screening Reagent is not reactive with these microorganisms: | ||
| Viruses | 32 | 31 | |
| Bacteria | 25 | 18 | |
| Chlamydiaspp. | 3 | 1 | |
| Characteristicand cell lines) | D3 Duet DFA Influenza A/Respiratory Virus Screening Kit | D3 Ultra DFA Respiratory VirusScreening & ID Kit | |
| Yeast | 1 | 0 | |
| Protozoan | 1 | 0 | |
| Cell lines | 17 | 17 |
{3}------------------------------------------------
·
{4}------------------------------------------------
D³ DUET DFA INFLUENZA A/RESPIRATORY VIRUS SCREFNING KIT Page 5 of 13
Non-Clinical Performance:
Staining patterns of the phycoerythrin-labeled influenza A virus MAbs on influenza A virus infected cells were similar to those of the Predicate device.
Precision/Reproducibility:
Assay precision, intra-assay variability and inter assay variability were assessed with a panel of proficiency-level antigen control slides. The panel consisted of slides spotted with cell preparations of the following:
-
- Low level influenza A (Victoria strain)
-
- Mid level influenza A (Victoria strain)
-
- Low level influenza A (Victoria strain) mixed with Mid level RSV (Washington strain)
-
- Mid level influenza A (Victoria strain) mixed with Low level RSV (Washington strain)
-
- Low level respiratory virus (either influenza virus B {Taiwan strain}, adenovirus type 1, Parainfluenza virus types 1, 2, or 3 (strains C35, Greer, C243 respectively). This panel member was rotated during the 5-days of testing so that cach virus is tested twice.
-
- Negative no infected cells present
The low level is estimated to contain between 4 to 10% infected cells per cell spot. The mid level is estimated to contain between 20 to 25% infected cells per cell spot. Both levels were below the level used in quality control slides. Each panel member was re-coded daily to prevent its identification. Each panel was stained twice per day for 5-days by three diffcrent laboratorics.
The following results were recorded for both the control slide and the panel slide:
-
- Presence or absence of Yellow-gold fluorescence.
-
- Percent of cells exhibiting Yellow-gold fluorescence
-
- Presence or absence of Green fluorescence
- Percent of cells exhibiting Green fluorescence
The combined data for negative specimens -- no infected cells present - from the three sites demonstrates that the R-PF labeled and FITC labeled MAbs reproducibly do not stain uninfected cells. No fluorescent cells were scen in 100% (60/60) of the wells lacking infected cells.
{5}------------------------------------------------
The combined data from the three sites demonstrates reproducible detection of influenza A virus by the R-PE labeled MAbs. The presence of influenza A virus infected cells was reported in 95.3% (143/150) of the wells in which the infected cells were expected:
| Influenza A virus detection Summary | ||||
|---|---|---|---|---|
| PositiveControl Slide | Low LevelSlide | Mid-LevelSlide | Low Level withMid-Level RSV | Mid-Level withLow Level RSV |
| 100% (30/30) | 100% (30/30) | 100% (30/30) | 83.3% (25/30) | 93.3% (28/30) |
The combined data demonstrates the reproducibility of the detection of respiratory syncytial virus by the FITC labeled MAbs. The presence of respiratory syncytial virus infected cells was reported in 100% (90/90) of the wells in which the infected cells were expected:
Respiratory syncytial virus detection Summary
| Positive Control Slide | Low Level Influenza Awith Mid-Level RSV | Mid-Level Influenza Awith Low Level RSV |
|---|---|---|
| 100% (30/30) | 100% (30/30) | 100% (30/30) |
The combined data demonstrates that the presence of R-PE fluorescent cells reproducibly does not interfere with the detection of respiratory syncytial virus by the FITC labeled MAbs. The presence of respiratory syncytial virus infected cells was reported in 100% (53/53) of the wells in which the R-PE stained infected cells were present:
Respiratory syncytial virus detection in the presence of R-PE positive cells Summary
| Low Level R-PE stained cells with Mid-Level RSV | Mid-Level R-PE stained cells with LowLevel RSV |
|---|---|
| 100% (25/25) | 100% (28/28) |
The combined data from all three sites demonstrates that the presence of R-PE in the stain reproducibly does not interfere with the FITC staining of other viruses. The presence of influenza B virus infected cells was reported in 100% (36/36) of the wells in which the infected cells were expected. The presence of adenovirus infected cells was reported in 100% (36/36) of the wells in which the infected cells were expected. The presence of parainfluenza virus type 1 virus infected cells was reported in 100% (36/36) of the wells in which the infected cells were expected. The presence of parainfluenza virus type 2 virus infected cells was reported in 100% (36/36) of the wells in which the infected cells were expected. The presence of parainfluenza virus type 3 virus infected cells was reported in 100% (36/36) of the wells in which the infected cells were expected.
| Respiratory virus detection in the presence of R-PE Summary | |||||
|---|---|---|---|---|---|
{6}------------------------------------------------
D³ DUET DFA INFLUENZA A/RESPIRATORY VIRUS SCREENING KIT Page 7 of 13
| AdenovirusControl Slide | Low LevelAdenovirus | Influenza BVirus ControlSlide | Low LevelInfluenza BVirus | Parainfluenzatype 1Control Slide | Low LevelParainfluenzatype 1 |
|---|---|---|---|---|---|
| 100% (30/30) | 100% (6/6) | 100% (30/30) | 100% (6/6) | 100% (30/30) | 100% (6/6) |
| Parainfluenzatype 2Control Slide | Low LevelParainfluenzatype 2 | Parainfluenzatype 3Control Slide | Low LevelParainfluenzatype 3 | ||
| 100% (30/30) | 100% (6/6) | 100% (30/30) | 100% (6/6) |
The reproducibility study data demonstrates that the presence of R-PE in the stain reproducibly does not interfere with the detection of the 5 respiratory viruses by their respective FITC labeled MAbs.
Analytical specificity
Results for analytical detection limit for the seven viruses detected by the D3 Duet were reported in numbers of fluorescent cells per cell monolayer. Each master stock virus preparation was diluted in a ten-fold manner. Four wells of a 96-well cell culture plate were inoculated with each dilution. The plates were centrifuged at 700 xg for 60 minutes, and then incubated at 35° to 37°C for 24hours. Four wells from each dilution were stained with the D3 Duet. Each well was then examined at 200x magnification and the number of fluorescent cells counted. The table below lists the virus identity and strain along with the fluorescent cell count.
| Analytical Sensitivity of D3 Duet compared with that ofD3 Ultra MAbs | |||
|---|---|---|---|
| (values are numbers of fluorescent cells per cellmonolayer) | |||
| Virus strain | VirusDilutions frommaster stock | Fluorescent staining cells/well | |
| D3 Duet | D3 Ultra | ||
| Influenza A virus(PR, VR-95 H1N1) | 1x10-5 | 1, 3, 2, 6 | 1, 3, 0, 5 |
| 1x10-6 | 1, 0, 1, 1 | 0, 0, 1, 0 | |
| 1x10-7 | 0, 0, 0, 0 | 0, 0, 0, 0 | |
| Influenza B virus(Hong Kong, VR-823) | 1x10-4 | 4, 1, 6, 2 | 0, 4, 3, 5 |
| 1x10-5 | 1, 0, 1, 1 | 0, 0, 2, 2 | |
| 1x10-6 | 0, 0, 0, 0 | 0, 0, 0, 0 | |
| Adenovirus (Type8, VR-8) | 1x10-6 | 1, 1, 3, 5 | 1, 3, 2, 4 |
| 1x10-7 | 0, 0, 0, 0 | 0, 0, 0, 0 | |
| RSV (Washington, | 1x10-2 | 1, 0, 3, 4 | 2, 3, 2, 0 |
{7}------------------------------------------------
| Analytical Sensitivity of D3 Duet compared with that ofD3 Ultra MAbs | |||
|---|---|---|---|
| (values are numbers of fluorescent cells per cell monolayer) | |||
| Virus strain | VirusDilutions frommaster stock | Fluorescent staining cells/well | |
| D3 Duet | D3 Ultra | ||
| VR-1401) | 1x10-3 | 0, 1, 1, 0 | 2, 1, 0, 0 |
| 1x10-4 | 0, 0, 0, 0 | 0, 0, 0, 0 | |
| Parainfluenza 1 (C-35, VR-94) | 1x10-4 | 7, 7, 6, 8 | 9, 8, 4, 6 |
| 1x10-5 | 2, 2, 3, 0 | 1, 0, 2, 1 | |
| 1x10-6 | 0, 0, 0, 0 | 0, 0, 0, 0 | |
| Parainfluenza 2(Greer, VR-92) | 1x10-4 | 4, 0, 3, 1 | 4, 3, 1, 2 |
| 1x10-5 | 0, 2, 0, 0 | 0, 1, 1, 1 | |
| 1x10-6 | 0, 0, 0, 0 | 0, 0, 0, 0 | |
| Parainfluenza 3 (C243, VR-93) | 1x10-6 | 3, 3, 0, 6 | 1, 1, 3, 5 |
| 1x10-7 | 1, 0, 1, 1 | 1, 1, 1, 0 | |
| 1x10-8 | 0, 0, 0, 0 | 0, 0, 0, 0 |
Analytical reactivity (inclusivity) of the D3 Duet was evaluated using 10 influenza A virus and 4 influenza B virus strains. Four wells of a 96-well cell culture plate were inoculated with each viral strain (diluted to less than 20-TCID50 per 0.2-mL inoculum). The plates were centrifuged at 700xg for 60 minutes, and then incubated at 35° to 37°C for 24-hours. Four wells from cach strain were stained with the D3 Duet, and each well was then examined at 200x magnification and the number of fluorescent cells counted. The table below lists the virus identity and strain along with the fluorescent cell count.
| Analytical Reactivity (inclusivity) of D³ Duetwith various influenza A virus andinfluenza B virus strains(values are numbers of fluorescent cellsper cell monolayer) | |
|---|---|
| Influenzastrain | Fluorescent staining cells/cellmonolayer |
| Influenza AWisconsin/56/2005 | 3, 2, 1, 0 |
| Influenza A WS,VR-1520 (H1N1) | 6, 6, 6, 4 |
| Influenza A HongKong, VR-544(H3N2) | 3, 4, 5, 5 |
| Influenza A NewJersey, VR-897 | 9, 12, 14, 15 |
{8}------------------------------------------------
| Analytical Reactivity (inclusivity) of D³ Duetwith various influenza A virus andinfluenza B virus strains(values are numbers of fluorescent cellsper cell monolayer) | |
|---|---|
| Influenzastrain | Fluorescent staining cells/cellmonolayer |
| Influenza A Victoria, VR-822 (H3N2) | 3, 3, 3, 5 |
| Influenza A PR, VR-95 (H1N1) | 3, 9, 9, 6 |
| Influenza A Port Chalmers, VR-810 (H3N2) | 6, 6, 9, 10 |
| Influenza A Aichi, VR-547 (H3N2) | 3, 7, 9, 11 |
| Influenza A Denver, VR-546 (H1N1) | 13, 14, 11, 10 |
| Influenza A Mal, VR-98 (H1N1) | 8, 3, 6, 4 |
| Influenza B GL/1739/54, VR-103 | 7, 6, 7, 7 |
| Influenza B Taiwan/2/62, VR-295 | 3, 1, 2, 5 |
| Influenza B Hong Kong/5/72, VR-823 | 3, 2, 0, 1 |
| Influenza B Maryland/1/59, VR-296 | 5, 6, 6, 8 |
Based on the data presented above, the assay can reliably detect influenza A virus and influenza B virus strains exhibiting both temporal and geographical diversity at viral levels near the limit of detection in cell culture. Analytical sensitivity of the phycoerythrin-labeled influenza A MAbs of the D3 Duet was determined, and compared to that of the fluorescein-labeled influenza A MAbs of the D' Ultra. Cell monolayers of R-Mix in 96-well plates were inoculated with prepared virus stock of influenza A virus, Victoria strain, VR-822 (H3N2), diluted to give a TCID50 of 1 per 0.2-mL inoculum. The plates were incubated at 37℃ for 24 hours. Monolayers were stained using the procedures in the D Ultra's labeling or the D Duet's draft labeling. The assay was performed four times. Results indicate that analytical sensitivities of the phycoerythrin-labeled and the fluorcscein-labeled influenza A MAbs are not statistically different, by a paired t-test".
DHI-Duct_FluARespi_Sec05_510k-Summary printed.doc
ª 50% tissue culture infectivity dose
6 Microsoft Office Excel, Microsoft Corporation
{9}------------------------------------------------
Clinical Performance:
Direct fresh specimens:
A study was performed prospectively at three sites with 1203 fresh specimens that were received for respiratory virus testing. Each specimen was evaluated by the D3 Duet DFA Influenza A/Respiratory Virus Screening Kit and a cleared DSFA device for the presence of influenza A, influenza B, respiratory syncytial virus, adenovirus, parainfluenza 1, parainfluenza 2 and parainfluenza 3 in cells derived from clinical specimens. A total of nineteen specimens were excluded from analysis due to a site deviations, duplicate specimen, insufficient cell numbers, or high background. These exclusions left 1184 specimen results for analysis.
The following tables detail the summary of the comparison of the D3 Duet and the cleared DSFA comparator assay, combined for study sites 1, 2, and 3:
| D3 Duet R-PE identification of influenza A virus positive specimens | |||
|---|---|---|---|
| Direct Specimen (1184 Specimens) | D3 Ultra Final Identification(influenza A virus) | ||
| Pos | Neg | ||
| D3 Duet R-PE(influenza A virus) | Pos | 99 | 0 |
| Neg | 1 | 1084 | |
| Positive Percent Agreement (PPA) | 99% (99/100) | ||
| 95% CI- PPA | 94.5, 99.8% | ||
| Negative Percent Agreement (NPA) | 100%(1084/1084) | ||
| 95% CI- NPA | 99.7, 100% |
| D3 Duet FITC detection of influenza B virus, respiratory syncytial virus, adenovirus, and parainfluenza virus types 1, 2, and 3 viruses | |||
|---|---|---|---|
| Direct Specimen (1184 Specimens) | D3 Ultra Final Identification | ||
| Pos | Neg | ||
| D3 Duel FITC Screen | Pos | 386 | 0 |
| Neg | 0 | 798* | |
| Positive Percent Agreement (PPA) | 100% (386/386) | ||
| 95% CI- PPA | 99.0,100% | ||
| Negative Percent Agreement (NPA) | 100% (798/798) | ||
| 95% CI- NPA | 99.5,100% |
{10}------------------------------------------------
| Virus Follow-up Identification of 386 D³ Duet FITC Positive Specimens for influenza Bvirus, respiratory syncytial virus, adenovirus, and parainfluenza virus types 1, 2, and 3viruses, using D³ Ultra Identification Reagents | ||||||
|---|---|---|---|---|---|---|
| Sensitivity | Specificity | 95% CI | ||||
| Virus | TP /(TP+FN) | percent | forSensitivity | TN/(TN+FP) | percent | forSpecificity |
| Influenza Bvirus | 11/11 | 100% | 74.12, 100 | 1173/1173 | 100% | 99.7, 100 |
| Adenovirus | 52/52 | 100% | 93.1, 100 | 1132/1132 | 100% | 99.7, 100 |
| Parainfluenzatype 1 | 4/4 | 100% | 51.0, 100 | 1180/1180 | 100% | 99.7, 100 |
| Parainfluenzatype 2 | 1/1 | 100% | 20.1, 100 | 1183/1183 | 100% | 99.7, 100 |
| Parainfluenzatype 3 | 19/19 | 100% | 83.2, 100 | 1165/1165 | 100% | 99.7, 100 |
| RespiratorySyncytial Virus | 299/299 | 100% | 98.7, 100 | 885/885 | 100% | 99.6, 100 |
D³ DUET DFA INFLUENZA A/RESPIRATORY VIRUS SCREENING KIT Page 11 of 13
The D2 Duet's ability to identify influenza A virus using phycoerythrin in direct specimens was compared to the D3 Ultra's ability using fluorescein. The positive percent agreement was 99% (95% CI range of 94.5% to 99.8%). The negative percent agreement was 100% (95% CI range of 99.7% to 100%). When the ability of the D3 Duet to detect the six other respiratory viruses using fluorescein in direct specimens was compared to the D3 Ultra's ability using fluorescein, the positive percent agreement was 100% (95% CI range of 99.0% to 100%). The negative percent agreement was 100% (95% CI range of 99.5% to 100%).
Specimen type distribution:
Tables below show the study results by the claimed specimen type. Results from sites 1, 2, and 3 have been combined.
| Influenza A virus by specimen type | ||||||
|---|---|---|---|---|---|---|
| Specimentype | PPA | NPA | ||||
| TP /(TP+FN) | percent | 95%CI forPPA | TN/(TN + FP) | percent | 95% CI forNPA | |
| NPA | 61/62 | 98.4% | 91.4, 99.7 | 525/525 | 100% | 99.3, 100 |
| NPS | 38/38 | 100% | 90.8, 100 | 501/501 | 100% | 99.2, 100 |
| D³ Duet FITC detection of influenza B virus, respiratory syncytial virus, adenovirus,and parainfluenza virus types 1, 2, and 3 viruses by specimen type | ||||||
| Specimentype | PPA | NPA | ||||
| TP /(TP+FN) | percent | 95%CI forPPA | TN/(TN+FP) | percent | 95% CI forNPA |
Influenza A virus by specimen type
{11}------------------------------------------------
D³ DUFT DFA INFLUENZA A/RESPIRATORY VIRUS SCREENING KIT Page 12 of 13
| NPA | 196/196 | 100% | 98.1, 100 | 391/391 | 100% | 99.0, 100 |
|---|---|---|---|---|---|---|
| NPS | 173/173 | 100% | 97.8, 100 | 366/366 | 100% | 99.0, 100 |
Cultured specimens:
To evaluate the performance of this device using cultured clinical specimens, a fourth study was performed with 298 frozen specimens to compare performance of the D' Duet DFA Influenza A/Respiratory Virus Screening Kit with that of the predicate for the presence of Influenza A, Influenza B, Respiratory Syncytial Virus, Adenovirus, Parainfluenza 1, Parainfluenza 2 and Parainfluenza 3 (Para 3) from cultured clinical specimens. At Study Site 4, 298 frozen specimens were processed for cell culture testing in accordance with the procedure in the Comparator product insert (same procedure for both Subject and Comparator devices) using R-Mix Too™ FreshCells™ in 48/24-fill multi-well plates. All specimens at study site 4 were derived from nasopharyngeal specimens. The results of this study are presented below. The table below shows the age distribution for individuals studied at site 4:
| Site 4 (culture) – Age Distribution | |
|---|---|
| 0 - 1 month | 5 |
| >1 month - 2 years | 130 |
| >2 - 12 years | 44 |
| >12 - 21 years | 28 |
| 22- 30 years | 19 |
| 31- 40 years | 20 |
| 41- 50 years | 10 |
| 51 - 60 years | 9 |
| 61-70 years | 8 |
| 71 - 80 years | 6 |
| 81-90 years | 8 |
| >90 years | 5 |
| Unknown age | 6 |
| Total | 298 |
The following tables detail the results of the cell culture study's comparison of D3 Duet's phycoerythrin-labeled MAbs identification of influenza A virus positive specimens, and D2 Duet's fluorescein-labeled MAbs detection of influenza B virus, respiratory syncytial virus, adenovirus, and parainfluenza virus types 1, 2, and 3 positive specimens.
| Study Site 4 (culture) - D3 Duet R-PE identification of influenza A virus positive specimens | |||
|---|---|---|---|
| Cell Culture (298 Specimens) | D3 Ultra Final Identification(influenza A virus) | ||
| Pos | Neg | ||
| D3 Duet R-PE(influenza A virus) | Pos | 67 | 0 |
| Neg | 0 | 231 | |
| Positive Percent Agreement (PPA) | 100% (67/67) |
DHI-Duct_FluARespi_Sec05 510k-Summary printed.doc
{12}------------------------------------------------
D² DUET DFA INFLUENZA A/RESPIRATORY VIRUS SCREENING KIT Page 13 of 13
| 050/21 | 94 6 100% | |
|---|---|---|
| Negative Percent Agreement (NPA) | 100% 1731/23 | |
| 95% CI- NPA100.014 | 100%ﺍﻟﻤﺴﺘﻘﻠﺔ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘ |
Study Site 4 (culture) - D3 Duet FITC detection of influenza B virus, respiratory syncytial virus, adenovirus, and parainfluenza virus types 1, 2, and 3 viruses Cell Culture (298 Specimens) D3 Ultra Final Identification Pos Neg Pos 72 0 D3 Duet FITC Screen Neg 0 226 100% (72/72) Positive Percent Agreement (PPA) 95% CI- PPA 95.0,100% Negative Percent Agreement (NPA) 100% (226/226) 95% CI- NPA 98.4,100%
A variety of viral respiratory pathogens were isolated. Virus identification of De Duet FITC Positive Specimens using D3 Ultra Identification Reagents yiclded the following isolates: influenza A virus [prevalence 22.5% (67/298)], influenza B virus [prevalence 6.7% (20/298)], respiratory syncytial virus [prevalence 11.1% (33/298)], adenovirus [prevalence 3.4% (10/298)], parainfluenza type 1 virus [prevalence 1.7% (5/298)], parainfluenza type 2 virus [prevalence 1.0% (3/298)], and parainfluenza type 3 virus [prevalence 3.0% (9/298)]. There were sixteen co-infections as follows: three influenza A virus + parainfluenza type 3 virus, one influenza A virus + parainfluenza type 1 virus, one influenza A virus + parainfluenza type 2 virus, two influenza A virus + respiratory syncytial virus, one influenza A virus + adenovirus, one influenza B virus + parainfluenza type 2 virus, one influenza B virus + parainfluenza type 3 virus, one influenza B virus + respiratory syncytial virus, one respiratory syncytial virus + parainfluenza type 1 virus, two respiratory syncytial virus + parainfluenza type 3 virus, one adenovirus + parainfluenza type 1 virus and one adenovirus + parainfluenza type 3 virus.
{13}------------------------------------------------
DFPARTMENT OF HEALTH & HUMAN SERVICES
Image /page/13/Picture/1 description: The image shows the seal of the Department of Health & Human Services (HHS). The seal is circular and contains the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. In the center of the seal is an eagle with its wings spread.
Food and Drug Administration 2098 Gaither Road Rockville MD 20850
Gail R. Goodrum Vice President of Regulatory Affairs Diagnostic Hybrids, Inc. 1055 East State Street Suite 100 Athens, Ohio 45701
DEC 2 3 2008
Re: K081746
Trade/Device Name: D3 Duet DFA Influenza A/Respiratory Virus Screening Kit Regulation Number: 21 CFR 866.3330 Regulation Name: Influenza virus serological reagents Regulatory Class: Class I Product Code: GNW Dated: November 26, 2008 Received: November 28, 2008
Dear Ms. Goodrum:
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 r ou may , the survisions 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 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
ﻟﻤﺆﺳﺴﺎ
{14}------------------------------------------------
Page 2 -
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 advice for your device on our labeling regulation (21 CFR Part 801), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at 240-276-0450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). For questions regarding postmarket surveillance, please contact CDRH's Office of Surveillance and Biometric's (OSB's) Division of Postmarket Surveillance at 240-276-3474. For questions regarding the reporting of device adverse events (Medical Device Reporting (MDR)), please contact the Division of Surveillance Systems at 240-276-3464. 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 (240) 276-3150 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.
Sincerely yours,
Sally attayna
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
{15}------------------------------------------------
D3 DUET DFA INFLUENZA A/RESPIRATORY VIRUS SCREENING KIT
Page 1 of 1
SECTION 04 - INDICATIONS FOR USE
510(k) Number (if known): K081746
Device Name: D3 Duet DFA Influenza A/Respiratory Virus Screening Kit
Indication for Use: The Diagnostic Hybrids, Inc. device, D3 Duet DFA Influenza A/Respiratory Virus Screening Kit, is intended for the qualitative detection and identification of influenza A, while screening for influenza B virus, respiratory syncytial 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. 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 attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
| Prescription Use | AND/OR | Over |
|---|---|---|
| X | ||
| (Part 21 CFR 801 Subpart D) | ( |
r-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 Device Evaluation (ODE)
Division Sign-Off
Office of In Vitro Diagnostic Device Evaluation and Safety
209 1746 510(k)
§ 866.3330 Influenza virus serological reagents.
(a)
Identification. Influenza virus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to influenza in serum. The identification aids in the diagnosis of influenza (flu) and provides epidemiological information on influenza. Influenza is an acute respiratory tract disease, which is often epidemic.(b)
Classification. Class I (general controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 866.9.