(169 days)
No
The device description details a manual immunofluorescence assay read by a human using a fluorescence microscope. There is no mention of automated image analysis or any computational methods that would suggest the use of AI/ML.
No
This device is a diagnostic tool intended for the qualitative detection and identification of viral antigens, not for treating or preventing disease.
Yes
The "Intended Use / Indications for Use" section explicitly states that the device "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...from patients with signs and symptoms of respiratory infection." This describes a diagnostic purpose.
No
The device is a diagnostic kit that includes physical reagents (antibodies, wash solution, mounting fluid) and requires a fluorescence microscope for analysis. It is not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states that the device is 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." This describes a test performed in vitro (outside the body) on biological specimens to provide information about a patient's health status (presence of viral antigens).
- Device Description: The description details a laboratory test using reagents, slides, and a fluorescence microscope to analyze biological samples (cells from clinical specimens or cell culture). This is characteristic of an in vitro diagnostic test.
- Performance Studies: The document includes performance studies evaluating the device's ability to detect the target analytes in clinical specimens, comparing it to a predicate device. This is a standard requirement for IVD devices seeking regulatory clearance.
- Predicate Device: The mention of a "Predicate Device(s)" with a K number (K061101, D3 Ultra DFA Respiratory Virus Screening & ID Kit) further confirms that this device is being evaluated within the regulatory framework for IVDs. K numbers are associated with FDA clearances for medical devices, including IVDs.
All these elements align with the definition and characteristics of an In Vitro Diagnostic device.
N/A
Intended Use / Indications for Use
The Diagnostic Hybrids, Inc. device, D3 Duet DFA RSV/Respiratory Virus Screening 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. 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.
Product codes
GNW, LKT
Device Description
The MAbs furnished with the D' Duet DFA RSV/Respiratory Virus Screening Kit (D3 Duet) are the same MAbs that are contained in the Diagnostic Hybrids, Inc. (DHI) device D' Ultra DFA Respiratory Virus Screening & ID Kit (D' Ultra) [510(k) number K061101. November 20, 2006]. The Diagnostic Hybrids, Inc. device, D3 Duet DFA RSV/Respiratory Virus Screening Kit, uses a blend of viral antigen-specific murine MAbs. MAbs for RSV are directly labeled with R-phycoerythrin (R-PE) for the rapid detection and identification of RSV. MAbs for influenza A virus, influenza B 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 RSV/Respiratory Virus Screening Reagent R-phycoerythrin-labeled murine MAbs directed against influenza A virus and a mixture of fluoresceinlabeled murine MAbs directed against influenza A. influenza B. adenovirus, and parainfluenza virus types 1, 2, and 3. The buffered, stabilized, aqueous solution also contains Evans Blue as a counter-stain and 0.1% sodium azide as preservative.
- · Normal Mouse Gamma Globulin DFA Reagent 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 syncytial 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 D' Duet DFA RSV/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 respiratory syncytial 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 red by the Evans Blue counter-stain. If only goldenyellow fluorescent cells are present the specimen can be reported as positive for respiratory syncytial virus antigen. If only apple-green fluorescent cells are present, the particular virus is identified using the individual reagents from the Dr UltraTM DFA Respiratory Virus Screening & ID Kit (D- 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 Dr Ultra on new, separate cell preparations.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Nasal, nasopharyngeal
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Not Found
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
A prospective study was performed at three sites with 1203 fresh clinical specimens (nasal and nasopharyngeal swabs and aspirates) for direct specimen analysis. 1187 specimens were included in the final analysis. The specimens were evaluated by the D3 Duet DFA Influenza A/Respiratory Virus Screening Kit and a cleared DSFA device (D3 Ultra DFA Respiratory Virus Screening & ID Kit) for the presence of respiratory syncytial virus, influenza A, influenza B, adenovirus, parainfluenza 1, parainfluenza 2, and parainfluenza 3.
A fourth study was performed with 298 frozen nasopharyngeal specimens to evaluate the performance of the D3 Duet DFA RSV/Respiratory Virus Screening Kit using cultured clinical specimens. The specimens were processed for cell culture testing using R-Mix TooTM FreshCells™ in 48/24-fill multi-well plates, in accordance with the procedure for the D3 Ultra DFA Respiratory Virus Screening & ID Kit.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Non-Clinical Performance:
- Staining Patterns: Staining patterns of the phycoerythrin-labeled respiratory syncytial virus MAbs on respiratory syncytial 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.
- Negative specimens (no infected cells present): No fluorescent cells were seen in 100% (60/60) of the wells lacking infected cells.
- Respiratory syncytial virus detection (R-PE labeled MAbs): Reproducible detection in 98% (147/150) of expected wells.
- Influenza A virus detection (FITC labeled MAbs): Reproducible detection in 96.7% (87/90) of expected wells.
- Influenza A virus detection in presence of R-PE positive cells: Reproducible detection in 94.7% (54/57) of wells where R-PE stained infected cells were present, indicating no interference.
- Detection of other viruses (Influenza B, Adenovirus, Parainfluenza types 1, 2, 3) in presence of R-PE: 100% detection for Influenza B (36/36), Adenovirus (36/36), Parainfluenza type 1 (36/36), Parainfluenza type 2 (36/36), and Parainfluenza type 3 (36/36), indicating no interference.
- Detection Limit (Analytical Sensitivity): Evaluated by counting fluorescent cells per cell monolayer after serial dilutions of virus master stock. Sensitivity was comparable between D3 Duet and D3 Ultra MAbs for Influenza A, Influenza B, Adenovirus, RSV, Parainfluenza 1, 2, and 3.
- Analytical Reactivity (Inclusivity): Evaluated using 10 influenza A virus and 4 influenza B virus strains. The assay reliably detected all tested strains at viral levels near the limit of detection in cell culture.
- Analytical Specificity (Cross-Reactivity Testing):
- Tested against 32 virus strains and 17 host culture cell types. No cross-reactivity observed.
- Tested against 25 bacterial strains, 1 yeast, 3 Chlamydia sp., and 1 protozoan. No cross-reactivity observed, except for Staphylococcus aureus, where staining appeared as small points of fluorescence, likely due to binding of protein A.
Clinical Performance:
-
Direct Fresh Specimens (N=1187): Compared D3 Duet to D3 Ultra.
- RSV detection (D3 Duet R-PE vs. D3 Ultra):
- Positive Percent Agreement (PPA): 100% (300/300) (95% CI: 97.8, 100%).
- Negative Percent Agreement (NPA): 100% (887/887) (95% CI: 99.6, 100%).
- Influenza A, B, Adenovirus, Parainfluenza 1, 2, 3 detection (D3 Duet FITC Screen vs. D3 Ultra):
- PPA: 100% (186/186) (95% CI: 98.0, 100%).
- NPA: 100% (1001/1001) (95% CI: 99.6, 100%).
- Virus Follow-up Identification (of 187 D3 Duet FITC Positive Specimens using D3 Ultra Identification Reagents):
- Influenza A virus: Sensitivity 100% (100/100), Specificity 100% (1087/1087).
- Influenza B virus: Sensitivity 100% (11/11), Specificity 100% (1176/1176).
- Adenovirus: Sensitivity 100% (52/52), Specificity 100% (1135/1135).
- Parainfluenza type 1: Sensitivity 100% (4/4), Specificity 100% (1183/1183).
- Parainfluenza type 2: Sensitivity 100% (1/1), Specificity 100% (1186/1186).
- Parainfluenza type 3: Sensitivity 100% (19/19), Specificity 100% (1168/1168).
- Specimen Type Distribution (NPA and NPS): PPA and NPA were 100% for both specimen types across all viruses.
- RSV detection (D3 Duet R-PE vs. D3 Ultra):
-
Cultured Specimens (N=298 from Site 4): Compared D3 Duet to D3 Ultra.
- RSV detection (D3 Duet R-PE vs. D3 Ultra):
- PPA: 100% (33/33) (95% CI: 89.5, 100%).
- NPA: 100% (265/265) (95% CI: 98.6, 100%).
- Influenza A, B, Adenovirus, Parainfluenza 1, 2, 3 detection (D3 Duet FITC Screen vs. D3 Ultra):
- PPA: 100% (104/104) (95% CI: 96.4, 100%).
- NPA: 100% (194/194) (95% CI: 98.1, 100%).
- RSV detection (D3 Duet R-PE vs. D3 Ultra):
Key results indicate 100% agreement (both PPA and NPA) between the D3 Duet and the predicate device D3 Ultra for the detection of respiratory syncytial virus and the screening of other respiratory viruses in both direct and cultured clinical specimens.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
- Positive Percent Agreement (PPA)
- Negative Percent Agreement (NPA)
- Sensitivity
- Specificity
Predicate Device(s)
K061101, D3 Ultra DFA Respiratory Virus Screening & ID Kit
Reference Device(s)
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 866.3480 Respiratory syncytial virus serological reagents.
(a)
Identification. Respiratory syncytial virus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to respiratory syncytial virus in serum. Additionally, some of these reagents consist of respiratory syncytial virus antisera conjugated with a fluorescent dye (immunofluorescent reagents) and used to identify respiratory syncytial viruses from clinical specimens or from tissue culture isolates derived from clinical specimens. The identification aids in the diagnosis of respiratory syncytial virus infections and provides epidemiological information on diseases caused by these viruses. Respiratory syncytial viruses cause a number of respiratory tract infections, including the common cold, pharyngitis, and infantile bronchopneumonia.(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 § 866.9.
0
D' DUET DFA RSV/RESPIRATORY VIRUS SCREENING KIT
SECTION 05, 510(K) SUMMARY
Applicant:
DIAGNOSTIC HYBRIDS, INC. 1055 East State Street Suite 100 Athens, OHIO 45701
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:
July 01, 2008
Device Name:
Trade name - D Duet DFA RSV/Respiratory Virus Screening Kit Common name - Fluorescent antibody test for detecting respiratory syncytial virus. and screening for other respiratory viruses Classification name - Respiratory Syncytial Virus, Antibody, Ifa Product Code - GNW Regulation - 21 CFR 866.3480, Class I, respiratory syncytial 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 RSV/Respiratory Virus Screening 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
Image /page/0/Picture/13 description: The image shows the number K081928 at the top. Below that is the logo for Diagnostic Hybrids. The logo includes the company name in bold text, followed by the tagline "Integrating Science and Humanity". To the right of the text is an image of a person with their arms raised, next to a DNA strand.
DEC 2 3 2008
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 MAbs furnished with the D' Duet DFA RSV/Respiratory Virus Screening Kit (D3 Duet) are the same MAbs that are contained in the Diagnostic Hybrids, Inc. (DHI) device D' Ultra DFA Respiratory Virus Screening & ID Kit (D' Ultra) [510(k) number K061101. November 20, 2006].
The Diagnostic Hybrids, Inc. device, D3 Duet DFA RSV/Respiratory Virus Screening Kit, uses a blend of viral antigen-specific murine MAbs. MAbs for RSV are directly labeled with R-phycoerythrin (R-PE) for the rapid detection and identification of RSV. MAbs for influenza A virus, influenza B 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 RSV/Respiratory Virus Screening Reagent R-phycoerythrin-labeled murine MAbs directed against influenza A virus and a mixture of fluoresceinlabeled murine MAbs directed against influenza A. influenza B. adenovirus, and parainfluenza virus types 1, 2, and 3. The buffered, stabilized, aqueous solution also contains Evans Blue as a counter-stain and 0.1% sodium azide as preservative.
- · Normal Mouse Gamma Globulin DFA Reagent 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 syncytial virus, adenovirus, and parainfluenza
2
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 D' Duet DFA RSV/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 respiratory syncytial 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 red by the Evans Blue counter-stain. If only goldenyellow fluorescent cells are present the specimen can be reported as positive for respiratory syncytial virus antigen. If only apple-green fluorescent cells are present, the particular virus is identified using the individual reagents from the Dr Ultra™ DFA Respiratory Virus Screening & ID Kit (D- 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 Dr Ultra on new, separate cell preparations.
Technological Characteristics:
The DHI device, D' Duet, has been compared directly to the DHI device, D' Ultra, as the legally marketed device. The technology used in both devices is based on a standard immunofluorescence assay technique utilizing either phycoerythrin- or fluorescein-labeled MAbs. A summary is provided in Table 5.1 below:
| TABLE 12.1: Technological Characteristics Comparison | |||
|---|---|---|---|
| Characteristic | D³ Duet DFA RSV/ Respiratory | ||
| Virus Screening Kit | D³ Ultra DFA Respiratory Virus | ||
| Screening & ID Kit | |||
| Monoclonal antibodies (MAbs) | The RSV/Respiratory Virus DFA | ||
| Screening Reagent | |||
| contains 12 MAbs to 6 different | |||
| respiratory viruses (influenza A virus, | |||
| influenza B virus, adenovirus, | |||
| parainfluenza virus type 1, | |||
| parainfluenza virus type 2, | |||
| parainfluenza virus type 3), plus 2 | |||
| MAbs to respiratory syncytial virus. | The Respiratory Virus DFA Screening | ||
| Reagent | |||
| contains 12 MAbs to 6 different | |||
| respiratory viruses (influenza A virus, | |||
| influenza B virus, adenovirus, | |||
| parainfluenza virus type 1, | |||
| parainfluenza virus type 2, | |||
| parainfluenza virus type 3), plus 2 | |||
| MAbs to respiratory syncytial virus. | |||
| TABLE 12.1: Technological Characteristics Comparison | D³ Duet DFA RSV/ Respiratory Virus Screening Kit | D³ Ultra DFA Respiratory Virus Screening & ID Kit | |
| Characteristic | Direct labeling, |
- using R-phycoerythrin (R-PE) to label the MAbs to respiratory syncytial virus antigens
- using fluorescein isothiocyanate (FITC) to label all other MAbs with fluorescein moiety | Direct labeling,
- using fluorescein isothiocyanate (FITC) to label all MAbs with fluorescein moiety | |
| Labeling method | Influenza A virus, influenza B virus, adenovirus, parainfluenza virus type 1, parainfluenza virus type 2, parainfluenza virus type 3 | Influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus, parainfluenza virus type 1, parainfluenza virus type 2, parainfluenza virus type 3 | |
| Fluorescein-labeled MAbs | Respiratory syncytial virus (Phycoerythrin-labeled influenza A virus MAbs stain with golden-yellow fluorescence) | None (Fluorescein-labeled respiratory syncytial virus MAbs stain with apple-green fluorescence) | |
| Phycoerythrin-labeled MAbs | Cell Fixative is the same for both devices: Acetone | | |
| Cell Fixative | | | |
| 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 nuclear staining is uniformly bright.
Respiratory Syncytial Virus: The fluorescence is cytoplasmic and punctate with small inclusions in the syncytia.
Parainfluenza 1, 2, 3: The fluorescence is cytoplasmic and punctate with irregular inclusions. Types 2 and 3 cause the formation of syncytia.
Adenovirus: The fluorescence is cytoplasmic and punctate or bright nuclear on both. | | |
| Analytical sensitivity, according to 96-well cell culture plates infected with Flu A diluted to give a TCID50 of 1 per 0.2-mL inoculum (reported as average of 4 runs) | There is no significant difference between the two devices for analytical sensitivity. | | |
| | 27.7 ± 1.7 culture positives out of 96 | 28.7 ± 1.3 culture positives out of 96 | |
| Analytical specificity (for influenza A virus strains; MAbs are reactive with all listed strains) | Mabs to respiratory syncytial virus were shown to be reactive with these virus strains: | | |
| | 3 Respiratory syncytial virus strains: Long, VR-26 Group A, Wash, VR-1401 Group B, 9320, VR-955 Group B | 3 Respiratory syncytial virus strains: Long, VR-26 Group A, Wash, VR-1401 Group B, 9320, VR-955 Group B | |
| | Device Screening Reagent is not reactive with these numbers of microorganisms.* | | |
| Analytical specificity (cross reactivity studies; various strains of microorganisms and cell lines) | Viruses | 32 | 31 |
| | Bacteria | 25 | 18 |
| | Chlamydia spp. | 3 | 1 |
| | Yeast | 1 | 0 |
| | Protozoan | 1 | 0 |
| | Cell lines | 17 | 17 |
| | * Lists of these microorganisms are provided in Section 18 of this 510(k) submission. | | |
3
4
Non-Clinical Performance:
Staining patterns of the phycoerythrin-labeled respiratory syncytial virus MAbs on respiratory syncytial 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 RSV (Washington strain)
-
- Mid level RSV (Washington 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, Grecr, C243 respectively). This panel member was rotated during the 5-days of testing so that each 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 pancl member was re-coded daily to prevent its identification. Each panel was stained twice per day for 5-days by three different laboratories.
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-PE labeled and FITC labeled MAbs reproducibly do not stain uninfected cells. No fluorescent cells were seen in 100% (60/60) of the wells lacking infected cells.
The combined data from the three sites demonstrates reproducible detection of respiratory syncytial virus by the R-PF labeled MAbs. The presence of respiratory syncytial virus infected cells was reported in 98% (147/150) of the wells in which the infected cells were expected:
| Respiratory syncytial virus detection Summary | ||||
|---|---|---|---|---|
| Positive | ||||
| Control Slide | Low Level | |||
| Slide | Mid-Level | |||
| Slide | Low Level with | |||
| Mid-Level | ||||
| Influenza A | Mid-Level with | |||
| Low Level | ||||
| Influenza A | ||||
| 100% (30/30) | 100% (30/30) | 100% (30/30) | 90% (27/30) | 100% (30/30) |
5
The combined data demonstrates the reproducibility of the detection of Influenza A virus by the FITC labeled MAbs. The presence of Influenza A virus infected cells was reported in 96.7% (87/90) of the wells in which the infected cells were expected:
| Influenza A virus detection Summary | ||
|---|---|---|
| Positive Control Slide | Low Level Influenza A | |
| with Mid-Level RSV | Mid-Level Influenza A | |
| with Low Level RSV | ||
| 100% (30/30) | 90% (27/30) | 100% (30/30) |
The combined data demonstrates that the presence of R-PE fluorescent cells does not interfere with the detection of influenza A virus by the FITC labeled MAbs in a reproducible manner. The presence of influenza A virus infected cells was reported in 94.7% (54/57) of the wells in which the R-PE stained infected cells were present:
| Influenza A virus detection in the presence of R-PE positive cells summary | |
|---|---|
| Low Level R-PE stained cells with Mid- | |
| Level influenza A virus | Mid-Level R-PE stained cells with Low |
| Level influenza A virus | |
| 100% (27/27) | 90% (27/30) |
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.
| Adenovirus
Control Slide | Low Level
Adenovirus | Influenza B
Virus Control
Slide | Low Level
Influenza B
Virus | Parainfluenza
type 1
Control Slide | Low Level
Parainfluenza
type 1 |
|------------------------------------------|--------------------------------------|------------------------------------------|--------------------------------------|------------------------------------------|--------------------------------------|
| 100% (30/30) | 100% (6/6) | 100% (30/30) | 100% (6/6) | 100% (30/30) | 100% (6/6) |
| Parainfluenza
type 2
Control Slide | Low Level
Parainfluenza
type 2 | Parainfluenza
type 3
Control Slide | Low Level
Parainfluenza
type 3 | | |
| 100% (30/30) | 100% (6/6) | 100% (30/30) | 100% (6/6) | | |
miratory virus detection in the precence of D. DE. Summary
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.
Detection limit:
Results for analytical detection limit for the seven viruses detected by the De 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 24-hours. 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 of | |||
|---|---|---|---|
| D3 Ultra MAbs | |||
| (values are numbers of fluorescent cells per cell monolayer) | |||
| Virus strain | Virus Dilutions from master stock | Fluorescent staining cells/well | |
| D3 Duet | D3 Ultra | ||
| Influenza A virus | |||
| (PR, VR-95 IIINI) | 1x10-5 | 4, 1, 5, 4 | 1, 3, 0, 5 |
| 1x10-6 | 1, 2, 0, 3 | 0, 0, 1, 0 | |
| 1x10-7 | 0, 0, 0, 0 | 0, 0, 0, 0 | |
| Influenza B virus | |||
| (Hong Kong, VR-823) | 1x10-4 | 3, 3, 4, 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 (Type 8, VR-8) | 1x10-6 | 1, 1, 3, 3 | 1, 3, 2, 4 |
| 1x10-7 | 0, 0, 0, 0 | 0, 0, 0, 0 | |
| RSV (Washington, VR-1401) | 1x10-2 | 1, 1, 3, 3 | 2, 3, 2, 0 |
| 1x10-3 | 2, 0, 0, 1 | 2, 1, 0, 0 | |
| 1x10-4 | 0, 0, 0, 0 | 0, 0, 0, 0 | |
| Parainfluenza 1 (C-35, VR-94) | 1x10-4 | 6, 5, 8, 6 | 9, 8, 4, 6 |
| 1x10-5 | 0, 2, 4, 2 | 1, 0, 2, 1 | |
| 1x10-6 | 0, 0, 0, 0 | 0, 0, 0, 0 | |
| Parainfluenza 2 | |||
| (Greer, VR-92) | 1x10-6 | 5, 4, 2, 1 | 4, 3, 1, 2 |
| 1x10-7 | 0, 0, 1, 0 | 0, 1, 1, 1 | |
| 1x10-8 | 0, 0, 0, 0 | 0, 0, 0, 0 | |
| Parainfluenza 3 (C 243, VR-93) | 1x10-6 | 1, 2, 0, 3 | 1, 1, 3, 5 |
| 1x10-7 | 1, 0, 1, 0 | 1, 1, 1, 0 |
7
| Analytical Sensitivity of D. Duet compared with that of
Dº Ultra MAbs
(values are numbers of fluorescent cells per cell
| monolayer) | |||||
|---|---|---|---|---|---|
| Virus | |||||
| Dilutions from | |||||
| master stock | Fluorescent staining cells/well | ||||
| Virus strain | D3 Duet | D³ Ultra | |||
| 1×10-8 | 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 each strain were stained with the D 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 Duet
on various influenza A virus and influenza
B virus strains (values are numbers of
| fluorescent cells per cell monolayer) | |
|---|---|
| Influenza strain | Fluorescent staining cells/cell |
| monolayer | |
| Influenza A WS, VR- | |
| 1520 (H1N1) | 10, 8, 7, 7 |
| Influenza A Hong | |
| Kong, VR-544 | |
| (H3N2) | 12, 11, 11, 12 |
| Influenza A New | |
| Jersey, VR-897 | |
| (H1N1) | 8, 11, 10, 14 |
| Influenza A Victoria, | |
| VR-822 (H3N2) | 7, 9, 10, 11 |
| Influenza A PR, VR- | |
| 95 (H1N1) | 6, 9, 8, 11 |
| Influenza A Port | |
| Chalmers, VR-810 | |
| (H3N2) | 8, 11, 15, 9 |
| Influenza A Aichi, | |
| VR-547 (H3N2) | 16, 15, 14, 13 |
| Influenza A Denver, | |
| VR-546 (H1N1) | 6, 9, 9, 8 |
| Influenza A Mal, VR- | |
| 98 (H1N1) | 16, 13, 11, 15 |
| Influenza A | |
| A/NWS/33, VR-219 | 12, 17, 15, 10 |
8
Analytical Reactivity (inclusivity) of D Duet on various influenza A virus and influenza B virus strains (values are numbers of fluorescent cells per cell monolayer)
| Influenza strain | Fluorescent staining cells/cell monolayer |
|---|---|
| (H1N1) | |
| Influenza B | |
| Russia/69, VR-790 | 13, 14, 12, 15 |
| Influenza B | |
| Mass/3/66, VR-523 | 12, 19, 14, 13 |
| Influenza B | |
| Hong Kong/5/72, | |
| VR-791 | 8, 8, 9, 11 |
| Influenza B | |
| Maryland/1/59, | |
| VR-296 | 16, 12, 13, 12 |
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 specificity:
Cross-Reactivity Testing
The D3 Duet RSV/Respiratory Virus Screening Kit was tested for cross-reactivity against a variety of cells and microorganisms. Stringent conditions for crossreactivity testing were achieved by using a high concentration of the D3 Duet DFA Influenza RSV/Respiratory Virus Screening Reagent and relatively high titers of microorganisms. The D3 Duet DFA RSV/Respiratory Virus Screening Reagent was prepared at 1.5X the concentration that is provided in the kit. No cross-reactivity was observed for 32 virus strains or for 17 host culture cell types. Twenty-five bacterial strains, one yeast, three Chlamydia sp. and one protozoan were evaluated for cross-reactivity, including Staphylococcus aureus, a protein-Aproducing bacterium. Staining of S. aureus appeared as small points of fluorescence.
Thirty-two virus strains were tested for cross reactivity. Depending on the particular virus, 71 to 1,400 TCID50 were inoculated into shell vial culture and incubated for 24 to 48 hours, to yield a 1+ to 3+ infection, processed and stained with the 1.5X DFA Reagent according to the procedure as detailed in this product insert. No cross reactivity was observed for the viruses listed below:
| Virus Strains Tested for Cross Reactivity with D3 Duet DFA RSV/Respiratory Virus Screening Reagent | |||||
|---|---|---|---|---|---|
| Organism | Strain or Type | Inoculum (TCID50) | Organism | Strain or Type | Inoculum (TCID50) |
| Parainfluenza 4a | M-25, VR-1378 | 1,400 | CMV | Towne, VR-977 | 430 |
| Parainfluenza 4b | CH19503, VR-377 | 1,400 | CMV | Davis, VR-807 | 430 |
9
| Virus Strains Tested for Cross Reactivity with D³ Duet DFA RSV/Respiratory Virus Screening Reagent | |||||
|---|---|---|---|---|---|
| Organism | Strain or Type | Inoculum | |||
| (TCID50) | Organism | Strain or Type | Inoculum | ||
| (TCID50) | |||||
| Metapneumovirus | Subgroup A1 | 1,400 | CMV | AD169, VR-538 | 430 |
| Metapneumovirus | Subgroup A2 | 1,400 | Varicella-zoster | Webster, VR-916 | 430 |
| Metapneumovirus | Subgroup B1 | 1,400 | Varicella-zoster | Ellen, VR-1367 | 430 |
| Metapneumovirus | Subgroup B2 | 1,400 | Rhinovirus 39 | 209 Picornavirus, | |
| VR-340 | 1,400 | ||||
| Coronavirus | OC43, VR-1558 | 1,400 | Rubeola | Commercially | |
| available slides | |||||
| stained.* | |||||
| Coronavirus | 229E, VR-740 | 1,400 | Mumps | Commercially | |
| available slides | |||||
| stained.* | |||||
| HSV-1 | 1F, VR-733 | 71 | Echovirus | Types 4, 6, 9, 11, | |
| 30, 34 | Commercially | ||||
| available slides | |||||
| stained.* | |||||
| HSV-1 | MacIntyre, VR-539 | 71 | Coxsackievirus | Types B1, B2, B3, | |
| B4, B5, B6 | Commercially | ||||
| available slides | |||||
| stained.* | |||||
| HSV-2 | MS, VR-540 | 71 | |||
| HSV-2 | Strain G, VR-734 | 71 |
Seventeen host culture cell types were tested for cross-reactivity. Cell cultures were prepared in shell vial format. Confluent monolayers were stained with the 1.5X preparation of the D' Duet DFA RSV/Respiratory Virus Screening Reagent according to the procedure as detailed in the product insert, and then examined for cross-reactivity. No cross-reactivity was observed for the following:
| | Cell lines Tested for Cross Reactivity with D³ Duet DFA Influenza
A/Respiratory Virus Screening Reagent | | |
|----------|------------------------------------------------------------------------------------------------------------|---------|-----------|
| A549 | monolayer | pCMK | cell spot |
| BGMK | monolayer | pRhMK | cell spot |
| HEp-2 | monolayer | RD | monolayer |
| LLC-MK2 | monolayer | RhMK II | cell spot |
| MDCK | monolayer | pRK | monolayer |
| MRC-5 | monolayer | R-Mix | monolayer |
| MRHF | monolayer | Vero | cell spot |
| MvlLu | monolayer | W1-38 | cell spot |
| NCI-H292 | monolayer | | |
Thirty microorganisms, including 25 bacterial and one yeast cultures, three Chlamydia sp. and one protozoan commercially available slides, were stained with the 1.5X DFA Reagent according to the procedure as detailed in the product insert, then examined for cross reactivity. Except for Staphylococcus aureus, which was cross reactive with the D Duet DFA Influenza A/Respiratory Virus Screening Reagent, all other microorganisms tested negative. Reactivity with Staphylococcus aureus is more than likely due to binding the protein A produced by Staphylococcus aureus. Concentrations for each bacterial organism cultured by DHI for cross reactivity testing were determined from suspensions of the bacteria in PBS by spectrophotometer according to McFarland standards of levels 1.0 and 2.0 (equaling approximately 3.0 x 106 and 6.0 x 100 CFU per mL). Slides
10
were prepared with spots of 0.01-mL of the suspensions to give either 3.0 x 104 or 6.0 x 104 per spot. At the same time, 1-mL of each suspension was plated on an appropriate agar dish for colony confirmation. According to the confirmation agar cultures, initial concentrations of the bacterial organisms in the study ranged from 6.4 x 104 to 2.9 x 10' CFU. Microorganisms tested are listed below.
| Microorganisms Tested for Cross Reactivity with D³ Duet DFA Influenza | |
|---|---|
| A/Respiratory Virus Screening Reagent | |
| BACTERIA | CFU TESTED |
| Acholeplasma laidlawii | ~6 x 107 |
| Acinetobacter calcoaceticus | 9.7 x 105 |
| Bordetella bronchiseptica | 1.7 x 105 |
| Bordetella pertussis | 4.6 x 106 |
| Corynebacterium diphtheriac | 2.5 x 106 |
| Escherichia coli | 2.6 x 105 |
| Gardnerella vaginalis | 5.0 x 105 |
| Haemophilis influenzae type A | 9.3 x 105 |
| Klebsiella pneumoniae | 6.4 x 106 |
| Legionella pneumophila | 6.5 x 104 |
| Moraxella cartarrhalis | 6.4 x 104 |
| Mycoplasma hominis | ~6 x 104 |
| Mycoplasma orale | ~6 x 104 |
| Mycoplasma pneumoniae | ~6 x 104 |
| Mycoplasma salivarium | ~6 x 107 |
| Neisseria gonorrhoeae | 1.3 x 106 |
| Proteus mirabilis | 2.1 x 106 |
| Pseudomonas aeruginosa | 1.0 x 107 |
| Salmonella enteriditis | 2.5 x 106 |
| Salmonella typhimurium | 1.8 x 106 |
| Staphylococcus aureus* | 1.0 x 107 |
| Streptococcus agalactiae | 9.6 x 106 |
| Streptococcus pneumoniae | 8.0 x 106 |
| Streptococcus pyogenes | 2.9 x 107 |
| Ureaplasma uralyticum | ~6 x 104 |
| Chlamydophila pneumoniae | Commercially available slides stained. |
| Chlamydophila psittaci | Commercially available slides stained. |
| Chlamydia trachomatis | Commercially available slides stained. |
| YEAST | |
| Candida glabrata | 8.7 x 106 |
| PROTOZOAN | |
| Trichomonas vaginalis | Commercially available slides stained. |
*Reactivity with Staphylococcus aureus is more than likely due to binding the protein A produced by Staphylococcus aureus.
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 D Duet DFA Influenza A/Respiratory Virus Screening Kit and a cleared DSFA device for the presence of respiratory syncytial virus, influenza A, influenza B, adenovirus, parainfluenza 1, parainfluenza 2 and parainfluenza 3 in cells derived from clinical specimens. Seventeen specimens were excluded from analysis due to a variety of reasons (site deviations, duplicate specimen, insufficient cell
11
numbers, or high background). These exclusions left 1187 specimen results for analysis.
and. John cleared DSFA comparator assay, combined for study sites 1, 2, and 3:
| D3 Duet R-PE identification of respiratory syncytial virus positive specimens | |||
|---|---|---|---|
| Direct Specimen (1187 Specimens) | D' Ultra Final Identification | ||
| (respiratory syncytial virus) | |||
| Pos | Neg | ||
| D3 Duet R-PE | Pos | 300 | 0 |
| (respiratory syncytial virus) | Neg | 0 | 887 |
| Positive Percent Agreement (PPA) | 100% | ||
| (300/300) | |||
| 95% CI-PPA | 97.8, 100% | ||
| Negative Percent Agreement (NPA) | 100% (887/887) | ||
| 95% CI- NPA | 99.6. 100% |
D3 Duet FITC detection of influenza A virus, influenza B virus, adenovirus, and parainfluenza virus types 1, 2, and 3
| D3 Ultra Final Identification | |||
|---|---|---|---|
| Direct Specimen (1187 Specimens) | D3 Duet FITC Screen | Pos | Neg |
| Pos | 187 | 0 | |
| Neg | 0 | 1000* | |
| Positive Percent Agreement (PPA) | 100% (186/186) | ||
| 95% CI- PPA | 98.0,100% | ||
| Negative Percent Agreement (NPA) | 100% (1001/1001) | ||
| 95% CI- NPA | 99.6,100% |
Virus Follow-up Identification of 187 D3 Duet FITC Positive Specimens for influenza A virus, influenza B virus, adenovirus, and parainfluenza virus types 1, 2, and 3 viruses, using D3 Ultra Identification Reagents
| Sensitivity | 95%CI | Specificity | 95% CI | |||
|---|---|---|---|---|---|---|
| Virus | TP / | |||||
| (TP+FN) | percent | for | ||||
| Sensitivity | TN/ | |||||
| (TN+FP) | percent | for | ||||
| Specificity | ||||||
| Influenza A | ||||||
| virus | 100/100 | 100% | 96.3, 100 | 1087/1087 | 100% | 99.7, 100 |
| Influenza B | ||||||
| virus | 11/11 | 100% | 74.1, 100 | 1176/1176 | 100% | 99.7, 100 |
12
Diagnostic Hybrids, Inc.
D3 DUET DFA RSV/RESPIRATORY VIRUS SCREENING KIT Section 5 - Page 13 of 15
| Adenovirus | 52/52 | 100% | 93.1, 100 | 1135/1135 | 100% | 99.7, 100 |
|---|---|---|---|---|---|---|
| Parainfluenza type 1 | 4/4 | 100% | 51.0, 100 | 1183/1183 | 100% | 99.7, 100 |
| Parainfluenza type 2 | 1/1 | 100% | 20.7, 100 | 1186/1186 | 100% | 99.7, 100 |
| Parainfluenza type 3 | 19/19 | 100% | 83.2, 100 | 1168/1168 | 100% | 99.7, 100 |
The D3 Duel's ability to identify respiratory syncytial virus using phycoerythrin in direct specimens was compared to the D3 Ultra's ability using fluorescein. The positive percent agreement was 100% (95% CI range of 98.7% to 100%). The negative percent agreement was 100% (95% CI range of 99.6% 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 97.8% to 100%). The negative percent agreement was 100% (95% CI range of 99.6% 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.
| Respiratory syncytial virus by specimen type Study Sites 1, 2, and 3 Combined | ||||||
|---|---|---|---|---|---|---|
| Specimen | ||||||
| type | PPA | 95%CI for | NPA | 95% CI | ||
| TP / | ||||||
| (TP+FN) | percent | PPA | TN/ (TN+FP) | percent | for NPA | |
| NPA | 155/155 | 100% | 97.6, 100 | 435/435 | 100% | 99.1, 100 |
| NPS | 132/132 | 100% | 97.2, 100 | 410/410 | 100% | 99.1, 100 |
D' Duet FITC detection of influenza A virus, influenza B virus, adenovirus, and parainfluenza virus types 1, 2, and 3 viruses by specimen type Study Sites 1, 2, and 3 Combined
| Specimen
| type | PPA | NPA | ||||
|---|---|---|---|---|---|---|
| TP / | ||||||
| (TP+FN) | percent | 95% CI | ||||
| for PPA | TN/ | |||||
| (TN+FP) | percent | 95% CI | ||||
| for NPA | ||||||
| NPA | 103/103 | 100% | 96.4, 100 | 484/484 | 100% | 99.2, 100 |
| NPS | 79/79 | 100% | 95.4, 100 | 460/460 | 100% | 99.2, 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 D3 Duet DFA RSV/Respiratory Virus Screening Kit with that of the predicate for the presence of respiratory syncytial virus, influenza A virus, influenza B virus, adenovirus, parainfluenza virus types 1, 2 and 3 from cultured
13
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 TooTM 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 respiratory syncytial virus specimens positive specimens, and D3 Duet's fluorescein-labeled MAbs detection of influenza A virus, influenza B virus, adenovirus, and parainfluenza virus types 1, 2, and 3 positive specimens.
| Cell Culture (298 Specimens) | | D³ Ultra Final Identification
(respiratory syncytial virus) | |
|-----------------------------------------------|-----|----------------------------------------------------------------|----------------|
| | | Pos | Neg |
| D³ Duet R-PE
(respiratory syncytial virus) | Pos | 33 | 0 |
| | Neg | 0 | 265 |
| Positive Percent Agreement (PPA) | | 100% (33/33) | |
| 95% CI- PPA | | 89.5, 100% | |
| Negative Percent Agreement (NPA) | | | 100% (265/265) |
| 95% CI- NPA | | | 98.6, 100% |
| Study Site 4(culture) - D3 Duet FITC detection of influenza A virus, influenza B virus,
| adenovirus, and parainfluenza virus types 1, 2, and 3 | |||
|---|---|---|---|
| Cell Culture (298 Specimens) | D3 Ultra Final Identification | ||
| Pos | Neg |
14
Diagnostic Hybrids, Inc.
D3 DUET DFA RSV/RESPIRATORY VIRUS SCREENING KIT Section 5 - Page 15 of 15
| Pos | 104 | 0 | |
|---|---|---|---|
| D3 Duet FITC Screen | Neg | 0 | 194 |
| Positive Percent Agreement (PPA) | 100% | ||
| (104/104) | |||
| 95% CI- PPA | 96.4,100% | ||
| Negative Percent Agreement (NPA) | 100% (194/194) | ||
| 95% CI- NPA | 98.1,100% |
A variety of viral respiratory pathogens were isolated. Virus identification of D3 Duet FITC Positive Specimens using D3 Ultra Identification Reagents yielded the following isolates: respiratory syncytial virus [prevalence 11.1% (33/298)], influenza A virus [prevalence 22.5% (67/298)], influenza B virus [prevalence 6.7% (20/298)], adenovirus [prevalence 3.4% (10/298)], parainfluenza type I 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.
15
Image /page/15/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized eagle with three bars forming its body and wings. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the eagle.
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
Re: K081928
Trade/Device Name: D3 Duet DFA RSV/Respiratory Virus Screening Kit Regulation Number: 21 CFR 866.3480 Regulation Name: Respiratory syncytial virus serological reagents Regulatory Class: Class I Product Code: LKT 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 general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.
DEC 2 3 2008
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 aft 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
16
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 attaym
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
17
SECTION 04, INDICATIONS FOR USE
510(k) Number (if known): K081928
Device Name: D3 Duet DFA RSV/Respiratory Virus Screening Kit
Indication for Use: The Diagnostic Hybrids, Inc. device, D3 Duet DFA RSV/Respiratory Virus Screening 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 attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
Prescription Use X AND/OR (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)
(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED
Concurrence of CDRH, Office of Device Evaluation (ODE)
Division Sign-Off
Office of In Vitro Diagnostic Device Evaluation and Safety
KO8 1928 SIG(K).