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K Number
K061101
Device Name
D3 ULTRA DFA RESPIRATORY VIRUS SCREENING & ID KIT
Manufacturer
DIAGNOSTIC HYBRIDS, INC.
Date Cleared
2006-11-20

(214 days)

Product Code
GNW
Regulation Number
866.3330
Type
Traditional
Panel
Microbiology
Reference & Predicate Devices
K022713
Predicate For
K081746,K081928,K091171,K092300,K092882,K093233,K093415
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP AuthorizedThirdpartyExpeditedreview
Intended Use

The Diagnostic Hybrids, Inc. D3 Ultra DFA (direct fluorescent antibody) RESPIRATORY VIRUS SCREENING & ID KIT is intended for the qualitative detection and identification of the Influenza A, Influenza B, Respiratory Syncytial Virus (RSV), Adenovirus, Parainfluenza 1, Parainfluenza 2 and Parainfluenza 3 virus in respiratory specimens, by either direct detection or cell culture method, by immunofluorescence using fluoresceinated monoclonal antibodies (MAbs). It is recommended that specimens found to be negative after examination of the direct specimen result be confirmed by cell culture. Negative results do not preclude respiratory virus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.

Device Description

The Diagnostic Hybrids, Inc. D3 Ultra DFA RESPIRATORY VIRUS SCREENING & ID KIT includes a DFA Screening Reagent that contains a blend of murine monoclonal antibodies (MAbs) directed against seven respiratory viruses (Influenza A. Influenza B. Respiratory Syncytial Virus, Adenovirus, Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3) plus seven separate DFA Reagents, each consisting of MAb blends directed against a single respiratory virus. The kit can be used for direct specimen or cell culture screening and final virus identification.

AI/ML Overview

Here's a breakdown of the acceptance criteria and study information for the Diagnostic Hybrids, Inc. D3 Ultra DFA Respiratory Virus Screening & ID Kit, based on the provided 510(k) summary:

Acceptance Criteria and Device Performance (Comparison of Subject Device vs. Predicate Device)

The study's acceptance criteria are implicitly defined by the demonstration of substantial equivalence to the predicate device. The performance is reported in terms of Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) with 95% Confidence Intervals (CI).

Test Type & Specimen StatusAgreement TypePredicate Result (Count)Subject Result (Count)Reported Performance (PPA/NPA)95% Confidence Interval
Direct Specimen Testing - Fresh Prospectively Collected Specimens (n=326)
Screen +PPA909495.5%89.0-98.2%
NegativeNPA23623298.3%95.7-99.3%
AdenovirusPPA1818100.0%82.4-100%
AdenovirusNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100.0%95.2-100%
Influenza APPA3232100.0%89.3-100%
Influenza ANPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100.0%94.2-100%
Influenza BPPA1819100.0%82.4-100%
Influenza BNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)98.7%92.9-99.8%
Parainfluenza 1PPA22100.0%34.2-100%
Parainfluenza 1NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100.0%96.0-100%
Parainfluenza 2PPA00--- (n=0)---
Parainfluenza 2NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100.0%96.1-100%
Parainfluenza 3PPA25100.0%34.2-100%
Parainfluenza 3NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)96.7%90.8-98.9%
RSVPPA1818100.0%82.4-100%
RSVNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100.0%95.2-100%
Direct Specimen Testing - Frozen Prospectively Collected Specimens (n=474)
Screen +PPA168168100%97.8-100%
NegativeNPA306306100%98.8-100%
AdenovirusPPA88100%63.1-100%
AdenovirusNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%97.7-100%
Influenza APPA8585100%95.7-100%
Influenza ANPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%95.6-100%
Influenza BPPA1919100%82.3-100%
Influenza BNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%97.6-100%
Parainfluenza 1PPA33100%38.3-100%
Parainfluenza 1NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%97.8-100%
Parainfluenza 2PPA33100%38.3-100%
Parainfluenza 2NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%97.8-100%
Parainfluenza 3PPA99100%70.1-100%
Parainfluenza 3NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%97.6-100%
RSVPPA5151100%93.0-100%
RSVNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%96.7-100%
Cell Culture Testing - Frozen Prospectively Collected Specimens (n=490)
Screen +PPA181181100%98.0-100%
NegativeNPA309309100%98.5-100%
AdenovirusPPA1313100%73.4-100%
AdenovirusNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%97.3-100%
Influenza APPA9393100%95.2-100%
Influenza ANPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%95.0-100%
Influenza BPPA2323100%83.1-100%
Influenza BNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%97.1-100%
Parainfluenza 1PPA66100%55.7-100%
Parainfluenza 1NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%97.4-100%
Parainfluenza 2PPA44100%45.4-100%
Parainfluenza 2NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%97.4-100%
Parainfluenza 3PPA99100%65.5-100%
Parainfluenza 3NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%96.6-100%
RSVPPA4949100%91.3-100%
RSVNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%96.6-100%
Study 3a-DS – Archival Parainfluenza Specimens (n=26)
Screen +PPA1718100%78.4-100%
NegativeNPA9888.9%54.3->99.9%
Parainfluenza 1PPA11100%16.8-100%
Parainfluenza 1NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%78.4-100%
Parainfluenza 2PPA55100%51.1-100%
Parainfluenza 2NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%73.4-100%
Parainfluenza 3PPA1112100%70.0-100%
Parainfluenza 3NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)85.7%46.7-99.5%
Study 3a-CC - Archival Parainfluenza Specimens (n=29)
Screen +PPA2121100%81.8-100%
NegativeNPA88100%62.8-100%
Parainfluenza 1PPA33100%38.3-100%
Parainfluenza 1NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%79.3-100%
Parainfluenza 2PPA55100%51.1-100%
Parainfluenza 2NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%77.3-100%
Parainfluenza 3PPA1313100%73.4-100%
Parainfluenza 3NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%62.8-100%
Study 3b-CC - Archival Clinical Isolates (n=81)
Screen +PPA8181100%94.6-100%
NegativeNPA00100% (n=0)97.3-100%
AdenovirusPPA1111100%70.0-100%
AdenovirusNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%93.8-100%
Influenza APPA1818100%79.3-100%
Influenza ANPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%93.1-100%
Influenza BPPA1717100%78.4-100%
Influenza BNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%93.2-100%
Parainfluenza 1PPA44100%45.4-100%
Parainfluenza 1NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%94.3-100%
Parainfluenza 2PPA11100%16.8-100%
Parainfluenza 2NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%94.5-100%
Parainfluenza 3PPA2626100%84.8-100%
Parainfluenza 3NPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%92.2-100%
RSVPPA55100%51.1-100%
RSVNPA(not directly given but inferred from overall negative)(not directly given but inferred from overall negative)100%94.2-100%

1. Sample sizes used for the test set and the data provenance:

  • Comparison Studies (Primary Clinical Evaluation):
    • Total specimens: 849 original specimens.
    • Direct Specimen (DS) Testing:
      • Fresh prospectively collected: 326 specimens.
      • Frozen prospectively collected: 474 specimens.
    • Cell Culture (CC) Method Testing: 520 specimens (subset of the 849). Of these, 490 were frozen prospectively collected.
    • Archived Parainfluenza Specimens (Study 3a-DS/CC):
      • DS: 26 specimens
      • CC: 29 specimens
    • Archived Clinical Isolates (Study 3b-CC): 81 clinical isolates.
  • Data Provenance:
    • Main study (849 specimens): All but 30 were prospectively collected during the 2005-2006 season. These 30 were archived as Parainfluenza-positive.
    • Archived Parainfluenza Specimens (Study 3a): Frozen original specimens previously determined to contain Parainfluenza (types 1, 2, or 3) during the 2006 "respiratory season," obtained from an additional laboratory.
    • Archived Clinical Isolates (Study 3b): Banked clinical isolates known to contain respiratory viruses from the 2005/2006 respiratory season.
    • Country of Origin: Not explicitly stated, but the submission is to the US FDA by "Diagnostic Hybrids, Inc. Athens, OHIO 45701", implying US-based studies.

2. Number of experts used to establish the ground truth for the test set and the qualifications of those experts:

  • The document implies that the "Predicate device" results acted as the reference or ground truth for the comparison studies. Therefore, the "experts" are the lab personnel who interpreted the predicate device results.
  • The number and specific qualifications of these experts are not explicitly stated in the provided text. The evaluation was conducted at three laboratory sites.

3. Adjudication method for the test set:

  • The document does not describe a specific adjudication method (e.g., 2+1, 3+1). It states that the "Subject" results were compared to "Predicate Results." This implies the predicate results were taken as the standard. In the case of Study 3a (Archived Parainfluenza), it mentions that "Original results reported by the laboratory were unknown to the study investigator," suggesting an independent comparison to pre-existing results rather than a real-time adjudication process.

4. If a multi-reader multi-case (MRMC) comparative effectiveness study was done, if so, what was the effect size of how much human readers improve with AI vs without AI assistance:

  • No, this section is not applicable. The device is an in vitro diagnostic (IVD) test kit for direct detection or cell culture by immunofluorescence using monoclonal antibodies. It is not an AI-assisted diagnostic tool for human readers. Therefore, an MRMC study comparing human readers with and without AI assistance was not performed.

5. If a standalone (i.e., algorithm only without human-in-the-loop performance) was done:

  • Yes, in a sense. The described performance is the "Subject device" (the test kit itself) compared against a "Predicate device" (another test kit). This represents the performance of the assay/reagent system in a laboratory setting, which is analogous to a "standalone" performance for such an IVD device, as it doesn't involve an AI algorithm or human interpretation outside of standard microscopy and reading of fluorescent signals.

6. The type of ground truth used:

  • The primary ground truth used for the comparison studies was the results obtained from a legally marketed predicate device (Diagnostic Hybrids, Inc. DFA Respiratory Virus Screening & ID Kit, K022713).
  • For the "Archived Parainfluenza Specimens" (Study 3a) and "Archived Clinical Isolates" (Study 3b), the ground truth was also based on previously determined results (implicitly from a predicate or similar method) for known positive specimens/isolates.

7. The sample size for the training set:

  • For the comparative effectiveness studies (clinical evaluations), there is no explicit mention of a separate "training set" for the device itself in the context of an algorithm or machine learning. The studies described are validation studies comparing the performance of the new device to a predicate. The device is a reagent-based test kit, not an algorithm that requires a training set in the typical AI sense.
  • However, during the development of the monoclonal antibodies (MAbs) and the kit, internal analytical studies would have been performed. The closest equivalent to "training" for such a device would be the extensive analytical testing described, such as detection limits and analytical specificity testing across various viral strains, host cell types, and bacterial cultures.

8. How the ground truth for the training set was established:

  • As mentioned above, the concept of a "training set" in the context of an algorithm is not directly applicable here.
  • For the analytical studies (detection limit, analytical specificity), the "ground truth" was established by:
    • Known viral stocks: Inoculating cell culture plates with specific viruses at known concentrations (e.g., "50 viral particles," "1 viral particle per every 2 wells," "250 virus/mL").
    • Known microorganisms and cell types: Testing against "64 virus strains (cultured and processed for staining)," "18 host culture cell types," and "18 bacterial cultures," where their identity was pre-established.

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Image /page/0/Picture/0 description: The image contains the logo for Diagnostic Hybrids. The logo features the company name in bold, capitalized letters, with the word "DIAGNOSTIC" stacked above "HYBRIDS". To the right of the text is a stylized graphic that resembles a DNA strand intertwined with a human figure. Above the logo, there is a handwritten code "KO6110".

510 (k) Summary

November 13, 2006

NUV 2 U 2006

1

A. 510(k) Number: K061101

B. Purpose for Submission: New device

C. Measurand:

Influenza A, Influenza B, Respiratory Syncytial Virus (RSV), Adenovirus, Parainfluenza 1, Parainfluenza 2 and Parainfluenza 3 virus in respiratory specimens

D. Type of Test:

Direct detection or cell culture method, by immunofluorescence using fluoresceinated monoclonal antibodies (MAbs)

E. Applicant:

Diagnostic Hybrids, Inc. 350 West State Street Athens, OHIO 45701 Tel. 740-593-1784 Fax. 740-597-1546 Contact person: Gail R. Goodrum

F. Proprietary and Established Names:

D3 Ultra DFA Respiratory Virus Screening & ID Kit Common Name: DFA (Direct Fluorescent Antibody) test kit for the identification of 7 respiratory viruses (Influenza A, Influenza B, Respiratory Syncytial Virus (RSV), Adenovirus, Parainfluenza 1, Parainfluenza 2 and Parainfluenza 3 virus) in patient specimens and cell cultures

G. Regulatory Information:

    1. Regulation section:
      866.3330 Influenza virus serological reagents
    1. Classification:
      Class I
    1. Product code: GNW
    1. Panel:

Microbiology (83)

H. Intended Use:

    1. Intended use(s):
      The Diagnostic Hybrids, Inc. D3 Ultra DFA (direct fluorescent antibody) RESPIRATORY VIRUS SCREENING & ID KIT is intended for the qualitative detection and identification of the Influenza A, Respiratory Syncytial Virus

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(RSV), Adenovirus, Parainfluenza 1, Parainfluenza 2 and Parainfluenza 3 virus in respiratory specimens. by either direct detection or cell culture method, by immunofluorescence using fluoresceinated monoclonal antibodies (MAbs). It is recommended that specimens found to be negative after examination of the direct specimen result be confirmed by cell culture. Negative results do not preclude respiratory virus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.

    1. Indication(s) for use:
      The Diagnostic Hybrids, Inc. D3 Ultra DFA (direct fluorescent antibody) RESPIRATORY VIRUS SCREENING & ID KIT is intended for the qualitative detection and identification of the Influenza A. Respiratory Syncytial Virus (RSV), Adenovirus, Parainfluenza 1, Parainfluenza 2 and Parainfluenza 3 virus in respiratory specimens, by either direct detection or cell culture method, by immunofluorescence using fluoresceinated monoclonal antibodies (MAbs). It is recommended that specimens found to be negative after examination of the direct specimen result be confirmed by cell culture. Negative results do not preclude respiratory virus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.
    1. Special conditions for use statement(s): For prescription use only
    1. Special instrument requirements: Fluorescence microscope with the correct filter combination for FITC (excitation peak = 490 nm, emission peak = 520nm).

I. Device Description:

The Diagnostic Hybrids, Inc. D3 Ultra DFA RESPIRATORY VIRUS SCREENING & ID KIT includes a DFA Screening Reagent that contains a blend of murine monoclonal antibodies (MAbs) directed against seven respiratory viruses (Influenza A. Influenza B. Respiratory Syncytial Virus, Adenovirus, Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3) plus seven separate DFA Reagents, each consisting of MAb blends directed against a single respiratory virus. The kit can be used for direct specimen or cell culture screening and final virus identification.

Kit Components:

  • Respiratory Virus DFA Screening Reagent a mixture of fluorescein labeled murine . monoclonal antibodies directed against respiratory viral antigens of Influenza A, Influenza B, Respiratory Syncytial Virus (RSV), Adenovirus, Parainfluenza 1, Parainfluenza 2 and Parainfluenza 3. The buffered, stabilized, aqueous solution contains Evan's Blue as a counter-stain and 0.1% sodium azide as preservative.
  • . Influenza A DFA Reagent - fluorescein labeled murine monoclonal antibodies directed against antigens produced by Influenza A virus infected cells. The buffered, stabilized, aqueous solution contains Evan's Blue as a counter-stain and 0.1% sodium azide as preservative.
  • Influenza B DFA Reagent fluorescein labeled murine monoclonal antibodies directed . against antigens produced by Influenza B virus infected cells. The buffered, stabilized, aqueous solution contains Evan's Blue as a counter-stain and 0.1% sodium azide as

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preservative.

  • RSV DFA Reagent fluorescein labeled murine monoclonal antibodies directed against . antigens produced by RSV infected cells. The buffered, stabilized, aqueous solution contains Evan's Blue as a counter-stain and 0.1% sodium azide as preservative.
  • . Adenovirus DFA Reagent - fluorescein labeled murine monoclonal antibodies directed against antigens produced by Adenovirus infected cclls. The buffered, stabilized, aqueous solution contains Evan's Blue as a counter-stain and 0.1% sodium azide as preservative.
  • Parainfluenza 1 DFA Reagent fluorescein labeled murine monoclonal antibodies . directed against antigens produced by Parainfluenza 1 infected cells. The buffered, stabilized, aqueous solution contains Evan's Blue as a counter-stain and 0.1% sodium azide as preservative.
  • Parainfluenza 2 DFA Reagent fluorescein labeled murine monoclonal antibodies . directed against antigens produced by Parainfluenza 2 infected cells. The buffered, stabilized, aqueous solution contains Evan's Blue as a counter-stain and 0.1% sodium azide as preservative.
  • . Parainfluenza 3 DFA Reagent - fluorescein labeled murine monoclonal antibodies directed against antigens produced by Parainfluenza 3 infected cells. The buffered, stabilized, aqueous solution contains Evan's 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 as to the virus infected cells present, i.e., Influenza A. Influenza B, Respiratory Syncytial Virus (RSV), Adenovirus, Parainfluenza 1, Parainfluenza 2 and Parainfluenza 3. The Negative well contains uninfected cells. Each slide is intended to be stained only one time.
  • Normal Mouse Gamma Globulin DFA Reagent a mixture of fluorescein labeled . murine gamma globulin that has been shown to be unreactive with any of the listed respiratory viruses. The buffered, stabilized, aqueous solution contains Evan's Blue as a counter-stain and 0.1% sodium azide as preservative.
  • Wash Solution Concentrate a 40X concentrate consisting of Tween 20 and 4% sodium . azide (after dilution to 1X in water, the concentration of sodium azide in the solution is 0.1%) in Phosphate Buffered Saline.
  • Mounting Fluid an aqueous, buffered, stabilized solution of glycerol and 0.1% sodium . azide.

J. Substantial Equivalence Information:

    1. Predicate device name(s): DFA Respiratory Virus Screening & ID Kit
    1. Predicate 510(k) number(s): K022713
    1. Comparison with predicate: The similarities to predicate device are in indicated use, operating principle, basic design, materials and formulation.

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Similarities
ItemDevicePredicate
Intended UseFor the qualitativedetection andidentification of therespiratory viruses,Influenza A, Influenza B,Respiratory SyncytialVirus (RSV), Adenovirus,Parainfluenza 1,Parainfluenza 2 andParainfluenza 3 virus byeither direct detection orcell culture methodFor the qualitativedetection andidentification of therespiratory viruses,Influenza A, Influenza B,Respiratory SyncytialVirus (RSV), Adenovirus,Parainfluenza 1,Parainfluenza 2 andParainfluenza 3 virus inrespiratory specimens,by either direct detectionor cell culture method
Basic principleDFA (Direct FluorescentAntibody) test -Immunofluorescenceusing fluoresceinatedmonoclonal antibodies(MAbs)DFA (Direct FluorescentAntibody) test -Immunofluorescenceusing fluoresceinatedmonoclonal antibodies(MAbs)
AntibodyBlend of murinemonoclonal antibodies(MAbs) directed againstseven respiratory virusesBlend of murinemonoclonal antibodies(MAbs) directed againstseven respiratory viruses
Instrumentation(required but notprovided)Fluorescencemicroscope with thecorrect filtercombination for FITC(excitation peak = 490nm, emission peak =520nm).Fluorescencemicroscope with thecorrect filtercombination for FITC(excitation peak = 490nm, emission peak =520nm).
Sample typeRespiratory specimensRespiratory specimens

Diagnostic Hybrids, Inc.

K. Standard/Guidance Document Referenced (if applicable): N/A

L. Test Principle:

The test kit uses viral antigen-specific murine monoclonal antibodies that are directly labeled with fluorescein for rapid detection and identification of respiratory viruses. The cells to be tested, either derived from a clinical specimen or cell culture, are fixed in acetone. The DFA Screening Reagent is added to the cells to determine the presence of viral antigens. After incubating at 35℃ to 37℃, the stained cells are rinsed 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. Virus infected cells will be stained with viral specific apple-green fluorescence when stained with the DFA

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Screening Reagent while uninfected cells will contain no fluorescence but will be stained red by the Evan's Blue counter-stain. If on examination of a direct stained specimen, no fluorescent-stained cells are found and all the cells stain red from the Evan's Blue, it is recommended that the specimen be cultured and stained using the DFA Screening Reagent. If fluorescent cells are seen, the particular virus is identified using the separate DFA Reagents on new, separate cell preparations. Cell preparations are fixed in acetone. The individual virus DFA Reagents are added to the cell preparations. After incubating at 35° to 37°C, the stained cells are rinsed with the diluted Wash Solution, a drop of the supplied Mounting Fluid is added and a coverslip is placed on the stained cells. The cells are examined using a fluorescence microscope for the presence of viral specific apple-green fluorescence, by which the unknown respiratory virus is identified.

Interpretation of results:

It is recommended that controls be examined first to ensure proper test performance before examination of the specimens. A positive reaction is one in which bright apple-green fluorescence is observed in the infected cells. Uninfected cells will stain dull red due to the Evan's Blue counter-stain included in the DFA Reagent.

Fluorescent staining pattern of respiratory virus infected cells:

The "typical" apple-green fluorescence staining pattern for each virus is as follows: 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.

Adenovirus: The fluorescence is cytoplasmic and punctate or bright nuclear or both. Parainfluenza 1, 2, 3: The fluorescence is cytoplasmic and punctate with irregular inclusions. Types 2 and 3 cause the formation of syncytia.

Co-infection with more than one infecting virus present in the specimen has been reported in a number of studies. The presence of multiple viruses is indicated when more than one well of the 8-well slide has fluorescent cells. The identification of the viruses is based on the individual virus DFA Reagents showing fluorescence. In such a case, it should be reported as " ... and ... detected by direct specimen testing." or " ... and ... isolated by cell culture."

Results From Direct Specimen Testing: The quality of the specimen with respect to the number of epithelial cells in the sample can be assessed by examining the different fields at a magnification of 200X. A satisfactory specimen should have at least 2 columnar epithelial cells per field. A negative result is indicated by the absence of fluorescence in a minimal sampling of 20 columnar epithelial cells. An inadequate sample is indicated by fewer than 20 columnar epithelial cells present in the sample, in which case the test is considered invalid. A new specimen should be obtained and tested or cell culture of the remaining specimen should be initiated.

A satisfactory specimen with no fluorescent cells found should be reported as "Presumptively negative, no Influenza A, Influenza B, Adenovirus, Respiratory Syncytial Virus, Parainfluenza 1, Parainfluenza 2, or Parainfluenza 3 detected by direct specimen testing". However, such negative results should be confirmed using cell culture. In the case culture yields a positive result, it should be reported as " ... isolated by cell culture", where '…' is the appropriate virus.

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Diagnostic Hybrids, Inc.

If fluorescent cells are found, continue with the Testing Procedure, staining with the individual virus DFA Reagents. The individual virus DFA Reagent that yields fluorescent cells represents the identification of the respiratory virus. In such a case, it should be reported as " ... detected by direct specimen testing', where ' ... ' is the appropriate virus, e.g. Influenza A, Influenza B, Adenovirus, Respiratory Syncytial Virus, Parainfluenza 1, Parainfluenza 2, or Parainfluenza 3.

Results from Culture Isolation / Confirmation: The entire cell spot or monolayer of cells must be examined for virus-infected, fluorescent cells. If no fluorescent cells are found, the results of testing of the specimen should be reported as, "No Influenza A, Influenza B, Adenovirus, Respiratory Syncytial Virus, Parainfluenza 1, Parainfluenza 2, or Parainfluenza 3 isolated by cell culture."

If fluorescent cells are found, continue with the Testing Procedure, staining with the individual virus DFA. The individual virus DFA Reagent that yields fluorescent cells represents the identification of the respiratory virus. In such a case, it should be reported as " ... isolated by cell culture", where ' ... ' is the appropriate virus, e.g. Influenza B, Respiratory Syncytial Virus, Parainfluenza 1, Parainfluenza 2, Parainfluenza 3, or Adenovirus.

M. Performance Characteristics (if/when applicable):

    1. Analytical performance:
    • a. Precision/Reproducibility: Not applicable
    • b. Linearity/assay reportable range: Not applicable
    • Traceability, Stability, Expected values (controls, calibrators, or methods): c. Not applicable
    • d. Detection limit:

The Predicate and Subject MAbs were compared by inoculating 96-well cell culture plates with the appropriate virus stock diluted to give ~ 50 viral particles for the entire plate or 1 viral particle per every 2 wells (i.e. 1 TCID50). The plates were incubated at 37° C for 24 hours and then stained with either the Subject Kit or the Predicate Kit. All plates were stained using the procedure in the product insert. This assay was performed 3 times with each virus and the results indicate no statistical difference from the Predicate kit by a paired t-test.

Each targeted respiratory virus was diluted to a value of 250 virus/mL and serial 2fold dilutions were then done down to a final value of 0.49 virus/mL. Each dilution of virus was inoculated into 6 shell vials of R-Mix, centrifuged at 700xg for 60 minutes and incubated at 35-37℃ for 48 hours. The Subject Kit or the Predicate Kit was used to stain 3 shell vials of each viral dilution according to the product insert. The sensitivity of both fluorescent antibody stains is equivalent, with ~ 0.5 - 1.0 PFU as the minimum viral dilution detected.

In order to determine if mixing the 15 various MAbs in the Screening Reagent affected their sensitivity, the analytical sensitivity of the subject screen reagent was compared to that of the subject individual reagents by inoculating 96-well cell culture plates with the appropriate virus stock diluted to give ~ 50 viral particles for

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the entire plate or 1 viral particle per every 2 wells. The plates were incubated at 37°C for 24 hours and then stained with cither the subject screening reagent or the subject individual reagents. All plates were stained using the procedure in the product insert. This assay was performed 3 times with each virus and the results indicate that the sensitivity of the screening reagent in the subject kit was not statistically different from that of the subject individual reagents using the paired test.

Analytical specificity:

The DFA Reagents were tested for cross-reactivity against a wide variety of cells and microorganisms. No cross-reactivity was observed for 64 virus strains (cultured and processed for staining) or for 18 host culture cell types. Eighteen (18) bacterial cultures were stained and examined for cross-reactivity, including Staphylococcus aureus, a protein-A-producing bacterium. Staining of S. aureus appeared as small points of fluorescence while all other bacterial cultures were negative. [Protein A will nonspecifically bind to the Fc portions of conjugated antibodies. Such binding can be distinguished from viral antigen binding on the basis of morphology, i.e., S. aureus-bound fluorescence appears as small (~1 micron diameter), bright dots. Stringent conditions for cross-reactivity testing were achieved by using high concentration DFAs and high titers of microorganisms. The DFAs (i.e. directly fluoresceinated monoclonal antibodies) were prepared at 1.5X the concentration that is provided in the kit. Each of the tested viruses was prepared as infected cell monolayers (250 infected cells inoculated into a shell vial culture and incubated for 24 to 48 hours, to yield a 3+ to 4+ infection), and processed and stained with the 1.5X DFAs according to the procedure detailed in the product insert. Bacterial strains were cultured, processed as suspensions, then spotted on microscope slides (vielding > 150 bacteria per 400X microscope field), then stained with the 1.5X DFAs according to the procedure in the product insert. Cell cultures were stained as confluent monolayers.

DFA Reagent (Results Positive (+) or Negative (-) for Reactivity)
OrganismStrainAdenoFlu AFlu BPara 1Para 2Para 3RSV
Type 1+------
Type 3+------
Type 5+------
Type 6+------
Type 7+------
AdenovirusType 10+------
Type 13+------
Type 14+------
Type 18+------
Type 31+------
Type 40+------
Type 41+------
Influenza AAichi (H3N2)-+-----
Mal (H1N1)-+-----
Hong Kong (H3N2)-+-----
Denver (H1N1)-+-----
Port Chalmers (H3N2)-+-----
Victoria (H3N2)-+-----
New Jersey (HswN1)-+-----
WS (H1N1)-+-----
PR (H1N1)+-1
Hong Kong기-+-
Maryland--+--l
Mass--+---
Influenza BTaiwan--+lﯿﮯ--
ভাこ+ー---
Russia--+----
Long------+
RSVWash------+
9320------+
Rhinovirus 39209 Picornavirus-------
C-35+
Parainfluenza 1------
Parainfluenza 2Greer--l-+--
Parainfluenza 3C 243"11-+1
Parainfluenza 4aM-25--I---l
Parainfluenza 4bCH18503-------
A1------
MetapneumovirusA2----l-
B3------
B4-------
CoronavirusOC43*-*יー
229E---י11
Herpes simplex virus1F-------
Type 1Macintyre----"・
Herpes simplex virusારક:-l--
Type 2Strain G-专-l-
Towneﯿﮯt---
CytomegalovirusDavis------
AD169י-י・--
Webster-----•-
Varicella-zosterEllen---I--1
9•ll-
11*【-
Echovirus301----l
34---
-Iי1ャ-
B1-l।--1-
B2---ਹ•--
Coxsackievirusвз----ーt
B4---l"--
BSーl1lーt-
BSー--י"+-
Mumps1----l
Rubeolaર-
Rhinovirus209 Picornavirus--",
Acholeplasma laidlawii---中--
Bordetella bronchiseptica---l-1
Bordetella pertussis--l-Iiﮯ
Chlamydia pneumoniae"----+
Clostridium diphtheriaeי-•---
Haemophillus influenzae type A-I-"---
Klebsiella pneumoniae---י---
Listeria pneumophila----י--
Moraxella cartarrhalis----ー
--
Mycobacterium avium----・--
Mycobacterium intracellulare-------
Mycoplasma hominis type 1t---"-I
Mycoplasma orale--"----
Mycoplasma pneumoniae--=1---
Mycoplasma salivarium-"--1--
Pseudomonas aeruginosa------
Streptococcus pneumoniae-・----l

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Diagnostic Hybrids, Inc.

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Diagnostic Hybrids, Inc.

Streptococcus pyogenes------
Ureaplasma urealyticum------
Cell cultures:
A549------
BGMK------
HEp-2------
LLC-MK2------
MDCK------
MRC-5------
MRHF------
Mv1Lu------
NCI-H292------
pCMK------
pRhMK------
pRK------
RD------
RhMK II------
R-Mix™------
R-Mix™ Too------
Vero------
WI-38------

f. Assay cut-off:

Not applicable

  1. Comparison studies:

a. Method comparison with predicate device:

The study included 849 original specimens evaluated by this product ("Subject" test) and a currently marketed DFA Screening & Identification Kit ("Predicate" test). All 849 specimens were studied by Direct Specimen (DS) testing with 22 of these specimens having insufficient cell numbers to be evaluated, and one other which could not be evaluated because it exhibited non-specific staining from the Normal Mouse Gamma Globulin DFA Reagent; 520 of the specimens also were studied by Cell Culture (CC) method with one specimen not evaluated because it produced a toxic cell culture monolayer. All but 30 of the specimens were prospectively collected during the 2005-2006 season; those 30 specimens had been archived as Parainfluenza-positive. In addition, a set of 81 clinical isolates were tested by CC methods only. The evaluations were conducted at three laboratory sites. Summary of results are shown in tables below:

Direct specimen testing – fresh prospectively collected specimens:
326 specimensNegativeScreen +AdenovirusInfluenza AInfluenza BParainfluenza 1Parainfluenza 2Parainfluenza 3Respiratory Syncytial Virus
Predicate Results:2369018321820218
Subject Results:2329418321920518
Positive Percent Agreement (PPA)95.5%100.0%100.0%100.0%100.0%--100.0%100.0%
95% Confidence Interval - PPA89.0-98.2%82.4-100%89.3-100%82.4-100%34.2-100%--34.2-100%82.4-100%
Negative Percent Agreement (NPA)98.3%100.0%100.0%98.7%100.0%100.0%96.7%100.0%
95% Confidence Interval - NPA95.7-99.3%95.2-100%94.2-100%92.9-99.8%96.0-100%96.1-100%90.8-98.9%95.2-100%

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Diagnostic Hybrids, Inc.
------------------------------
Direct specimen testing - frozen prospectively collected specimens:
474 specimensNegativeScreen +AdenovirusInfluenza AInfluenza BParainfluenza 1Parainfluenza 2Parainfluenza 3Respiratory Syncytial Virus
Predicate Results:3061688851933951
Subject Results:3061688851933951
PPA100%100%100%100%100%100%100%100%
95% CI - PPA97.8-100%63.1-100%95.7-100%82.3-100%38.3-100%38.3-100%70.1-100%93.0-100%
NPA100%100%100%100%100%100%100%100%
95% CI - NPA98.8-100%97.7-100%95.6-100%97.6-100%97.8-100%97.8-100%97.6-100%96.7-100%

Cell culture testing - frozen prospectively collected spec

490 specimensNegativeScreen +AdenovirusInfluenza AInfluenza BParainfluenza 1Parainfluenza 2Parainfluenza 3RespiratorySyncytial Virus
Predicate Results:30918113932364949
Subject Results:30918113932364949
PPA100%100%100%100%100%100%100%100%
95% CI – PPA98.0-100%73.4-100%95.2-100%83.1-100%55.7-100%45.4-100%65.5-100%91.3-100%
NPA100%100%100%100%100%100%100%100%
95% CI – NPA98.5 –100%97.3-100%95.0-100%97.1-100%97.4-100%97.4-100%96.6-100%96.6-100%

Specimens and culture isolates used in these studies came from nasopharyngeal (NP) aspirates, washes, swabs, bronchial alveolar lavages (BAL) and/or racheal aspirates.

  • b. Matrix comparison:
    n/a

    1. Clinical studies:
    • Clinical Sensitivity: ા. Not applicable.
    • b. Clinical specificity: Not applicable.
    • Other clinical supportive data (when a. and b. are not applicable): C.

Non-prospective archival specimens: Due to relative low prevalence of Parainfluenza infections in populations of respiratory specimens, few prospectively collected specimens were reactive with the Parainfluenza DFA Reagents. Frozen original specimens previously determined to contain Parainfluenza (types 1, 2, or 3) during the 2006 "respiratory season" were obtained from an additional laboratory, and were tested in an internal reference laboratory using the Subject and Predicate Tests by Wirect Specimen method (Study 3a-DS; see table below). The same specimens were tested by Cell Culture method (Study 3a-CC, see table below). Original results reported by the laboratory were unknown to the study investigator. Although the study desired by a

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Diagnostic Hybrids, Inc.

selection bias, this study offers further analytical information on the assay's ability to detect Parainfluenza viruses.

Study 3a-DS – Direct Specimen Results
26 specimensNegativeScreen +AdenovirusInfluenza AInfluenza BParainfluenza 1Parainfluenza 2Parainfluenza 3RespiratorySyncytialVirus
Predicate Results91700015110
Subject Results:81800015120
PPA100%------100%100%100%--
95% CI – PPA78.4% -100%------16.8% - 100%51.1% - 100%70.0% - 100%--
NPA88.9%100%100%100%100%100%85.7%100%
95% CI - NPA54.3% ->99.9%79.3% -100%79.3% -100%79.3% -100%78.4% - 100%73.4% - 100%46.7% -99.5%79.3% -100%
Study 3a-CC - Cell Culture Results
29 specimensNegativeScreen +AdenovirusInfluenza AInfluenza BParainfluenza 1Parainfluenza 2Parainfluenza 3Respiratory Syncytial Virus
Predicate Results82100035130
Subject Results:82100035130
PPA100%------100%100%100%--
95% CI - PPA81.8% - 100%------38.3% - 100%51.1% - 100%73.4% - 100%--
NPA100%100%100%100%100%100%100%100%
95% CI - NPA62.8% - 100%81.8% - 100%81.8% - 100%81.8% - 100%79.3% - 100%77.3% - 100%62.8% - 100%81.8% - 100%

Non-prospective archival clinical isolates: A study was conducted using a collection of banked clinical isolates known to contain respiratory viruses that had been frozen from the 2005/2006 respiratory season. These specimens were selected because they were previously shown to contain at least one of the seven virus analytes detacted by the Subject Test.

Study 3b-CC - Cell Culture Results
81 specimensNegativeScreen +AdenovirusInfluenza AInfluenza BParainfluenza 1Parainfluenza 2Parainfluenza 3RespiratorySyncytialVirus
PredicateResults:08111181741265
Subject Results:08111181741265
PPA100%100%100%100%100%100%100%100%
95% CI - PPA94.6%- 100%70.0% -100%79.3% -100%78.4% -100%45.4% - 100%16.8% - 100%84.8% - 100%51.1% -100%
NPA100%100%100%100%100%100%100%100%
95% CI - NPA97.3%- 100%93.8% -100%93.1% -100%93.2% -100%94.3% - 100%94.5% - 10092.2% - 100%94.2% -100%

4. Clinical cut-off:

Not applicable

    1. Expected values/Reference range: Respiratory virus infections are often seasonal, with Influenza typically extending from

{11}------------------------------------------------

November to April in the northern hemisphere, and Adenovirus infections occurring more often during late winter to early summer. RSV is usually a seasonal (winter and early spring) infection as well, with epidemics lasting up to 5 months, while outbreaks caused by parainfluenza viruses may occur throughout a year.

The clinical studies were comprised of respiratory specimens collected during the winter to carly spring months of 2005/2006. Prevalence of the respiratory viruses within the population of specimens that were prospectively collected and tested fresh is noted in the table below:

Expected ValuesAdenovirusInfluenza AInfluenza BParainfluenza 1Parainfluenza 2Parainfluenza 3RespiratorySyncytial Virus
Fresh Specimens(n = 326)18321920518
Prevalence5.5%9.8%5.8%0.6%01.5%5.5%

The following table summarizes the participant age demographics according to test results for a population of 326 fresh specimens, prospectively collected and evaluated for performance using the predicate assay:

Virus:Age *:AdenovirusInfluenza AInfluenza BParainfluenza 1Parainfluenza 2Parainfluenza 3RespiratorySyncytial VirusNegative
Totals18321820218236
<1m00000021
1m to 2y894102880
2y to12y876000142
12y to18y11500008
18y to 21y00100002
>21y0121000178
Not reported131100625

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/12/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized eagle with its wings forming a caduceus-like symbol. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES. USA" are arranged in a circular pattern around the eagle.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

NOV 2 0 2006

Ms. Gail R. Goodrum Vice President, Regulatory and Quality Affairs Diagnostic Hybrids, Inc. 350 West State Street Athens, OH 45701

Re: K061101

Trade/Device Name: D Ultra DFA Respiratory Virus Screening & ID Kit Regulation Number: 21 CFR 866.3330 Regulation Name: Influenza virus serological reagents Regulatory Class: Class I Product Code: GNW Dated: October 10, 2006 Received: October 11, 2006

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.

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).

{13}------------------------------------------------

This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (240)276-0450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 443-6597 or at its Internet address http://www.fda.gov/cdrh/dsma/dsmamain.html.

Sincerely yours.

Sally, atton

Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known): K061101

Device Name:__________________________________________________________________________________________________________________________________________________________________

Indications For Use:

The Diagnostic Hybrids, Inc.'s D3 Ultra DFA (direct fluorescent antibody) RESPIRATORY VIRUS SCREENING & ID KIT is intended for the qualitative detection and identification of the Influenza A, Influenza B, Respiratory Syncytial Virus (RSV), Adenovirus, Parainfluenza 1, Parainfluenza 2 and Parainfluenza 3 virus in respiratory specimens, by either direct detection or cell culture method, by immunofluorescence using fluoresceinated monoclonal antibodies (MAbs). It is recommended that specimens found to be negative after examination of the direct specimen result be confirmed by cell culture. Negative results do not preclude respiratory virus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.

Prescription Use (Part 21 CFR 801 Subpart D)

AND/OR

Over-The-Counter Use (21 CFR 807 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)

Division Sign-Off

Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K061101

Page 1 of 1

§ 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.