K Number
K093415
Date Cleared
2009-12-23

(51 days)

Product Code
Regulation Number
866.3400
Reference & Predicate Devices
Predicate For
N/A
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP AuthorizedThirdpartyExpeditedreview
Intended Use

The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit is intended for the qualitative identification of adenovirus and to screen for the presence of parainfluenza virus types 1, 2, and 3 in nasal and nasopharyngeal swabs and aspirates/washes specimens from patients with signs and symptoms of respiratory infection by direct detection of immunofluorescence using monoclonal antibodies (MAbs).

It is recommended that specimens found to be negative for parainfluenza virus and adenovirus after examination of the direct specimen result be confirmed by cell culture. Negative results do not preclude parainfluenza virus and adenovirus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.

Device Description

The D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit (D3 FastPoint PIV/ADV Kit) uses a blend (called an "L-DFA Reagent") of viral antigen-specific murine monoclonal antibodies that are directly labeled with either R-phycoerythin (PE) (parainfluenza virus types 1, 2 and 3) or fluorescein isothiocyanate (FITC) (adenovirus) for the qualitative identification of adenovirus and to screen for the presence of parainfluenza virus types 1, 2, and 3.

The cells to be tested are derived from respiratory specimens from patients with signs and symptoms of respiratory infection. The cells are permeabilized and stained concurrently in a liquid suspension format with the L-DFA Reagent. After incubating at 35℃ to 37℃ for 5-minutes, the stained cell suspensions are rinsed with 1X PBS. The rinsed cells are pelleted by centrifugation and then re-suspended with the Resuspension Buffer and loaded onto a specimen slide well. The cells are examined using a fluorescence microscope. Cells infected with parainfluenza virus types 1, 2 and 3 will exhibit golden-yellow fluorescence. Cells infected with adenovirus will exhibit apple green fluorescence due to the FITC. Non-infected cells will exhibit red fluorescence due to the Evans Blue counter-stain. Nuclei of intact cells will exhibit orange-red fluorescence due to the propidium iodide.

AI/ML Overview

Here's an analysis of the provided document regarding the acceptance criteria and study for the D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit:

1. Table of Acceptance Criteria and Reported Device Performance:

The document doesn't explicitly state "acceptance criteria" in a numerical target format (e.g., "sensitivity must be >90%"). Instead, it presents the achieved performance metrics in comparison to a composite comparator method (FDA-cleared device + viral culture). For the purpose of this table, I'll extract the reported performance from the clinical study, which implicitly serves as the demonstration of meeting acceptable clinical performance for substantial equivalence.

Acceptance Criteria (Implied by Study Results & Predicate Equivalence) and Reported Device Performance

Performance MetricAdenovirus (NP Wash/Aspirate)Parainfluenza Virus (NP Wash/Aspirate)Adenovirus (NP Swab)Parainfluenza Virus (NP Swab)
Sensitivity92.3% (95% CI: 64.0-99.8%)92.0% (95% CI: 74.0-99.0%)100% (95% CI: N/A - due to low prevalence)92.9% (95% CI: 66.1-99.8%)
Specificity100% (95% CI: 99.4-100%)99.3% (95% CI: 98.3-99.8%)100% (95% CI: 99.5-100%)100% (95% CI: 99.4-100%)
Reproducibility (Total Agreement)100% (All sites, for both Adenovirus and hPIV-1 in various combinations)100% (All sites, for both Adenovirus and hPIV-1 in various combinations)100% (All sites, for both Adenovirus and hPIV-1 in various combinations)100% (All sites, for both Adenovirus and hPIV-1 in various combinations)
Limit of Detection (LOD)100 infected cells/mL100 infected cells/mL (hPIV-1), 25 infected cells/mL (hPIV-2), 50 infected cells/mL (hPIV-3)Not applicable (analytical study)Not applicable (analytical study)
Analytical Reactivity (Inclusivity)Positive detection for 10 adenovirus strainsPositive detection for 3 hPIV strainsNot applicable (analytical study)Not applicable (analytical study)

2. Sample Size and Data Provenance for the Test Set:

  • Test Set (Clinical Performance Study):

    • Total Specimen Sample Size: 1519 specimens (across all age groups and specimen types).
    • NP Wash/Aspirate (Combined Sites 1, 2, 3):
      • Parainfluenza Virus: 628 specimens
      • Adenovirus: 632 specimens
    • NP Swab (Combined Sites 3, 4):
      • Parainfluenza Virus: 682 specimens
      • Adenovirus: 681 specimens
    • Data Provenance: Prospective, collected from 4 geographically diverse U.S. clinical laboratories during the 2009 respiratory virus season (January-March 2009). The specimens were "excess, remnants of respiratory specimens that were prospectively collected from symptomatic individuals suspected of respiratory infection" and were de-identified.
  • Test Set (Reproducibility Study):

    • Sample Size: A reproducibility panel consisting of 5 randomized panel members, tested daily in two separate runs for 5 days by four different laboratories (40 total runs). Each panel member contained defined levels of infected or non-infected cells.
    • Data Provenance: Not specified beyond being conducted by "four different laboratories." This would be part of a controlled laboratory study, not clinical specimens.

3. Number of Experts and Qualifications for Ground Truth (Clinical Test Set):

  • The document does not explicitly state the number of experts (e.g., medical doctors, virologists) used to establish the ground truth for the clinical test set.
  • Qualifications of Experts (Implied): The ground truth was established using a "composite comparator method" which included:
    • Direct Specimen Fluorescent Antibody (DSFA) test with an FDA-cleared device. This implies that the interpretation of the comparator DSFA test would have been performed by qualified laboratory personnel trained in using that specific FDA-cleared device.
    • Viral culture confirmation. This would have been performed by trained microbiologists or virologists capable of performing and interpreting viral cultures.

4. Adjudication Method for the Test Set (Clinical Study):

  • Adjudication Method: The ground truth was established using a composite comparator method.
    • "True" positive was defined as any sample that either tested positive by the comparator DSFA test or viral culture.
    • "True" negative was defined as any sample that tested negative by both the comparator DSFA test and viral culture.
    • This is a form of adjudicated reference standard, where agreement between multiple methods (or sequential application of methods, as implied by "negatives followed by culture") establishes the "truth." This method is commonly used when a single perfect gold standard is not available or practical.

5. Multi Reader Multi Case (MRMC) Comparative Effectiveness Study:

  • No, an MRMC comparative effectiveness study was not explicitly described. The study focused on the standalone performance of the D3 FastPoint kit against a composite comparator. There is no mention of human readers using the D3 FastPoint kit with and without AI assistance (as would be the case for a typical MRMC study involving AI) nor an effect size for human reader improvement with AI.

6. Standalone (Algorithm Only) Performance Study:

  • Yes, a standalone study was performed. The clinical performance section directly assesses the D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit's performance (device only, without human-in-the-loop assistance beyond the interpretation of the kit's results) against a composite comparator method. The sensitivity and specificity values reported in Tables 5.6, 5.7, 5.8, and 5.9 represent this standalone performance.

7. Type of Ground Truth Used (Clinical Study):

  • Composite Comparator Method: The ground truth for the clinical study was established using a composite comparator method. This method combined:
    • Performance of an existing FDA-cleared DSFA device.
    • Viral culture confirmation for all specimens negative by the initial comparator DSFA.
    • This combines elements of an established diagnostic method (FDA-cleared device) with a traditional gold standard for viral identification (viral culture).

8. Sample Size for the Training Set:

  • The document does not report on a training set sample size. This is because the device described is a diagnostic kit (reagents and controls for immunofluorescence), not an AI/machine learning algorithm that typically requires a distinct training phase. Performance is evaluated through analytical and clinical studies, not by training a model.

9. How Ground Truth for the Training Set Was Established:

  • Not applicable, as there is no training set described for an AI/machine learning model. The kit itself is the "algorithm" in a sense, and its performance is validated through analytical studies (reproducibility, LOD, inclusivity) and clinical studies against established comparator methods.

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Diagnostic Hybrids, Inc.

K093415

D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit

10/30/2009 DEC 2 3 2009 Page 1 of 13

Section 05, 510(k) Summary

Applicant:

DIAGNOSTIC HYBRIDS, INC. 1055 East State Street Suite 100 Athens, OHIO 45701

Contact Information:

Ronald H. Lollar, Senior Director Product Realization, Management, and Marketing 1055 East State Street Suite 100 Athens, Ohio 45701 740-589-3300 - Corporate number 740-589-3373 - Desk phone 740-593-8437 - Fax lollar@dhiusa.com

Date of preparation of 510(k) summary:

October 30, 2009

Device Name:

Trade name - D2 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit Common name - D3 FastPoint Parainfluenza Virus/Adenovirus Identification Kit Classification name - Parainfluenza virus serological reagents, Adenovirus serological reagents Product Code - GQS, GNY Regulation - 21 CFR 866.3400, 21 CFR 866.3020 Regulatory Class - Class I Panel Microbiology (83)

Legally marketed devices to which equivalence is claimed:

D3 Ultra DFA Respiratory Virus Screening & ID Kit (K061101)

Intended Use: The Diagnostic Hybrids, Inc. D3 Ultra DFA (direct fluorescent antibody) Respiratory Virus Screening & ID Kit (D3 Ultra) is intended for the qualitative detection and identification of the influenza A, influenza B, respiratory syncytial virus (RSV), adenovirus, parainfluenza 1, parainfluenza 2 and parainfluenza 3 virus in respiratory specimens, by either direct detection or cell culture method, by immunofluorescence using fluoresceinated monoclonal antibodies (MAbs). It is recommended that

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specimens found to be negative after examination of the direct specimen result be confirmed by cell culture. Negative results do not preclude respiratory virus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.

  • Performance characteristics for influenza A were established when . influenza A/H3 and A/H1 were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary.
  • If infection with a novel influenza A virus is suspected based on current . clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL3+ facility is available to receive and culture specimens.

D3 Duet DFA RSV/Respiratory Virus Screening Kit (K081928)

Intended Use: The Diagnostic Hybrids, Inc. device, D3 Duet DFA RSV/Respiratory Virus Screening Kit (D5 Duet RSV Kit), is intended for the qualitative detection and identification of respiratory syncytial virus, while screening for influenza A virus, influenza B virus, adenovirus, and parainfluenza virus types 1, 2 and 3 viral antigens, in nasal and nasopharyngeal swabs and aspirates or in cell culture. The assay detects viral antigens by immunofluorescence using monoclonal antibodies (MAbs), from patients with signs and symptoms of respiratory infection.

It is recommended that specimens found to be negative after examination of the direct specimen result be confirmed by cell culture. Negative results do not preclude influenza virus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.

Performance characteristics for influenza A virus detection and identification were established when influenza A (H3N2) and influenza A (H1N1) were the predominant influenza A strains circulating in the United States. Performance characteristics for influenza A virus detection and identification were established when influenza A H3N2 and influenza A H1N1 were the predominant influenza A strains circulating in the United States. When other influenza A viruses are emerging, performance characteristics may vary. If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to a state or local health department for testing. Viral

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culture should not be attempted in these cases unless a BSL3+ facility is available to receive and culture specimens.

Device Description:

The D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit (D3 FastPoint PIV/ADV Kit) uses a blend (called an "L-DFA Reagent") of viral antigenspecific murine monoclonal antibodies that are directly labeled with either Rphycoerythin (PE) (parainfluenza virus types 1, 2 and 3) or fluorescein isothiocyanate (FITC) (adenovirus) for the qualitative identification of adenovirus and to screen for the presence of parainfluenza virus types 1, 2, and 3.

Kit Components:

  • D3 FastPoint L-DFA PIV/Adenovirus Reagent, 4.0-mL. One dropper bottle 1. containing a mixture of PE-labeled murine monoclonal antibodies directed against parainfluenza virus types 1, 2, or 3 antigens and FITC-labeled murine monoclonal antibodies directed against adenovirus antigens. The buffered, stabilized, aqueous solution contains Evans Blue and propidium iodide as counter-stains and 0.1% sodium azide as preservative.
  • 40X PBS Concentrate, 25-mL. One bottle of 40X PBS concentrate containing 2. 4% sodium azide (0.1% sodium azide after dilution to 1X using de-mineralized water).
  • Re-suspension Buffer, 6.0-mL. One bottle of a buffered glycerol solution and 3. 0.1% sodium azide.
  • D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Antigen Control Slides, 4. 5-slides. Five individually packaged control slides containing 2 wells with cell culture-derived positive and negative control cells. Each positive well contains cells infected with either parainfluenza virus type 3 or adenovirus. The negative wells contain non-infected cells. Each slide is intended to be stained only one . time.
  • D3 FastPoint L-DFA Specimen Slides and Coverslips, 50-slides with 5. coverslips. Fifty pack of 3-well specimen slides.

The cells to be tested are derived from respiratory specimens from patients with signs and symptoms of respiratory infection. The cells are permeabilized and stained concurrently in a liquid suspension format with the L-DFA Reagent. After incubating at 35℃ to 37℃ for 5-minutes, the stained cell suspensions are rinsed with 1X PBS. The rinsed cells are pelleted by centrifugation and then re-suspended with the Resuspension Buffer and loaded onto a specimen slide well. The cells are examined using a fluorescence microscope. Cells infected with parainfluenza virus types 1, 2 and 3 will exhibit golden-yellow fluorescence. Cells infected with adenovirus will exhibit apple

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green fluorescence due to the FITC. Non-infected cells will exhibit red fluorescence due to the Evans Blue counter-stain. Nuclei of intact cells will exhibit orange-red fluorescence due to the propidium iodide.

Intended Use:

The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit is intended for the qualitative identification of adenovirus and to screen for the presence of parainfluenza virus types 1, 2, and 3 in nasal and nasopharyngeal swabs and aspirates/washes specimens from patients with signs and symptoms of respiratory infection by direct detection of immunofluorescence using monoclonal antibodies (MAbs).

It is recommended that specimens found to be negative for parainfluenza virus and adenovirus after examination of the direct specimen result be confirmed by cell culture. Negative results do not preclude parainfluenza virus and adenovirus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.

TABLE 5.1: Characteristics of the D³ FastPoint PIV/ADV Kit are compared to those of thefollowing Diagnostic Hybrids (DHI) predicate devices
CharacteristicsD³ FastPoint PIV/ADV Kit(Subject Device)D³ Ultra Kit510(k) #K061101D³ Duet RSV Kit510(k) # K081928
Intended UseThe Diagnostic Hybrids, Inc. device, D³ FastPoint L-DFAParainfluenza Virus/Adenovirus IdentificationKit is intended for thequalitative identification ofadenovirus and to screen forthe presence of parainfluenzavirus types 1, 2, and 3 innasal and nasopharyngealswabs and aspirates/washesspecimens from patients withsigns and symptoms ofrespiratory infection bydirect detection ofimmunofluorescence usingmonoclonal antibodies(MAbs).It is recommended thatspecimens found to benegative for parainfluenzavirus and adenovirus afterexamination of the directspecimen result be confirmedThe DiagnosticHybrids, Inc. D³Ultra™ DFA (directfluorescentantibody)Respiratory VirusScreening & ID Kitis intended for thequalitative detectionand identification ofthe influenza A,influenza B,respiratory syncytialvirus (RSV),adenovirus,parainfluenza 1,parainfluenza 2 andparainfluenza 3virus in respiratoryspecimens, by eitherdirect detection orcell culture method,byimmunofluorescenceusing monoclonalantibodies (MAbs).The DiagnosticHybrids, Inc. deviceD³ Duet DFARSV/RespiratoryVirus Screening Kit,is intended for thequalitative detectionand identification ofrespiratory syncytialvirus, whilescreening forinfluenza A virus,influenza B virus,adenovirus, andparainfluenza virustypes 1, 2 and 3 viralantigens, in nasaland nasopharyngealswabs and aspiratesor in cell culture.The assay detectsviral antigens byimmunofluorescenceusing monoclonalantibodies (MAbs).
TABLE 5.1:Characteristics of the D³ FastPoint PIV/ADV Kit are compared to those of the following Diagnostic Hybrids (DHI) predicate devices
CharacteristicsD³ FastPoint PIV/ADV Kit(Subject Device)D³ Ultra Kit510(k) #K061101D³ Duet RSV Kit510(k) # K081928
by cell culture. Negativeresults do not precludeparainfluenza virus andadenovirus infection andshould not be used as thesole basis for diagnosis,treatment or othermanagement decisions.antibodies (MAbs).It is recommendedthat specimensfound to be negativeafter examination ofthe direct specimenresult be confirmedby cell culture.Negative results donot precluderespiratory virusinfection and shouldnot be used as thesole basis fordiagnosis, treatmentor othermanagementdecisions.from patients withsigns and symptomsof respiratoryinfection.It is recommendedthat specimensfound to be negativeafter examination ofthe direct specimenresult be confirmedby cell culture.Negative results donot precludeinfluenza virusinfection and shouldnot be used as thesole basis fordiagnosis, treatmentor othermanagementdecisions.
Target Virusesadenovirus, parainfluenzavirus type 1, parainfluenzavirus type 2, parainfluenzavirus type 3influenza A virus,influenza B virus,respiratorysyncytial virus,adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 3influenza A virus,influenza B virus,respiratorysyncytial virus,adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 3
Monoclonal antibodies(MAbs)The D³ FastPoint L-DFAPIV/Adenovirus Reagentcontains 9 MAbs toadenovirus (3) andparainfluenza virus (6)The RespiratoryVirus DFAScreening Reagentcontains 15 MAbs to7 differentrespiratory viruses(influenza A virus,influenza B virus,respiratory syncytialvirus, adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 3)TheRSV/RespiratoryVirus DFAScreening Reagentcontains 15 MAbs to7 differentrespiratory viruses(influenza A virus,influenza B virus,adenovirus,parainfluenza virustype 1, parainfluenzavirus type 2,parainfluenza virustype 3), plus 2 MAbs
TABLE 5.1: Characteristics of the D³ FastPoint PIV/ADV Kit are compared to those of thefollowing Diagnostic Hybrids (DHI) predicate devices
CharacteristicsD³ FastPoint PIV/ADV Kit(Subject Device)D³ Ultra Kit510(k) #K061101D³ Duet RSV Kit510(k) # K081928syncytial virus.
Labeling methodDirect labeling- using R-Phycoerythrin (R-PE) to label the MAbs toparainfluenza virus types 1,2, and 3.- using fluoresceinisothiocyanate (FITC) tolabel the MAbs toadenovirus.Direct labeling- using fluoresceinisothiocyanate(FITC) to label allMAbs withfluorescein.Direct labeling- using R-Phycoerythrin (R-PE) to label theMAbs to respiratorysyncytial virus.- using fluoresceinisothiocyanate(FITC) to label allother MAbs withfluorescein.
R-Phycoerythrin-labeledMAbsparainfluenza virus types 1,2, and 3Nonerespiratory syncytialvirus
Fluorescein-labeled MAbsadenovirusinfluenza A virus,influenza B virus,respiratory syncytialvirus, adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 3influenza A virus,influenza B virus,adenovirus,parainfluenza virustype 1, parainfluenzavirus type 2,parainfluenza virustype 3
Cell FixativeProprietary Non-Acetonebased systemAcetoneAcetone
Cell Counter-stainPropidium Iodide, EvansBlueEvans BlueEvans Blue
Performance characteristics
Staining patternsParainfluenza 1, 2, 3: Thefluorescence is cytoplasmic.Cells appear round.Adenovirus: Thefluorescence is cytoplasmicor bright nuclear or both.Cells appear round.Negative: Cells fluorescered due to the Evans Bluecounter-stain.Nuclei: Cell Nucleifluoresce orange-red due tothe Propidium Iodidecounter-stain.Influenza A and B:The fluorescence iscytoplasmic, nuclearor both.Cytoplasmicstaining is oftenpunctate with largeinclusions whilenuclear staining isuniformly bright.RespiratorySyncytial Virus:The fluorescence iscytoplasmic andpunctate with smallinclusions in thesyncytia.Influenza A andB: Thefluorescence iscytoplasmic,nuclear or both.Cytoplasmicstaining is oftenpunctate with largeinclusions whilenuclear staining isuniformly bright.RespiratorySyncytial Virus:The fluorescenceis cytoplasmic andpunctate withsmall inclusions inthe syncytia.
TABLE 5.1: Characteristics of the D³ FastPoint PIV/ADV Kit are compared to those of thefollowing Diagnostic Hybrids (DHI) predicate devices
CharacteristicsD³ FastPoint PIV/ADV Kit(Subject Device)D³ Ultra Kit510(k) #K061101D³ Duet RSV Kit510(k) # K081928
3: The fluorescenceis cytoplasmic andpunctate withirregular inclusions.Types 2 and 3 causethe formation ofsyncytia.Adenovirus: Thefluorescence iscytoplasmic andpunctate or brightnuclear or both.Negative: Cellsfluoresce red due tothe Evans Bluecounter-stain.Parainfluenza 1,2, 3: Thefluorescence iscytoplasmic andpunctate withirregularinclusions. Types2 and 3 cause theformation ofsyncytia.Adenovirus: Thefluorescence iscytoplasmic andpunctate or brightnuclear or both.Negative: Cellsfluoresce red dueto the Evans Bluecounter-stain.
Analyticalspecificity(cross-reactivitystudies; variousstrains ofmicroorganisms and cell lines)VirusesDevice Reagents are not reactive with these numbers of microorganisms
Viruses593132
Bacteria221825
Chlamydiaspp.113
Yeast101
Protozoan001
Cell linesN/A1717

Technological Characteristics, Compared to Predicate Device:

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" Diagnostic Hybrids, Inc.

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Analytical Performance:

Precision/Reproducibility:

Assay precision, intra-assay variability and inter assay variability were assessed with a reproducibility panel consisting of 5 randomized panel members.

The hPIV/adenovirus panel consisted of the following

  • a. Low level parainfluenza virus type 1 (C-35 strain) infected cells.
  • b. Low level adenovirus (ATCC type 1) infected cells.
  • Low level parainfluenza virus type 1 (C-35 strain) infected cells mixed c. with mid level adenovirus (ATCC type 1) infected cells.
  • d. Low adenovirus (ATCC type 1) infected cells mixed with mid level parainfluenza virus type 1 (C-35 strain) infected cells.
  • Mid level non-infected (negative) cells. e.

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The low level is estimated to contain between 4 to 10% infected cells in the sample. The mid level is estimated to contain between 20 to 25% infected cells in the sample. Each sample contains 2.5 x 102 to 3.5 x 102 total cells.

Each panel was tested daily in two separate runs for 5-days by four different laboratories (40 total runs). The following results were recorded:

  • a. Presence or absence of golden-yellow fluorescence.
  • b. Percent of cells exhibiting golden-vellow fluorescence.
  • c. Presence or absence of apple-green fluorescence.
  • d. Percent of cells exhibiting apple-green fluorescence.

For the D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit, the combined data from the four Study Sites demonstrated reproducible detection of parainfluenza virus type 1 (hPIV-1) by the R-PE labeled MAbs and reproducible detection of adenovirus by the FITC-labeled MAbs. The presence of hPIV-1 infected cells was reported in 100% (120/120) of the wells in which the infected cells were expected. The presence of adenovirus infected cells was reported in 100% (120/120) of the wells in which the infected cells were expected. The absence of infected cells was reported in 100% (40/40) of the wells in which infected cells were not present. The total percent agreement for the D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit was 100% (280/280):

TABLE 5.2: Reproducibility Study Results using the L-DFA Reagent
SitePanelMemberNegativeAdenovirusLow LevelhPIV LowLevelMixed InfectionMixed Infection
ConcentrationNoinfectedcells4 to 10%infectedcells4 to 10%infectedcellsAdenovirusMid Level
Site1AgreementwithExpected result10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)70/70(100%)
Site2AgreementwithExpected result10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)70/70(100%)
Site3AgreementwithExpected result10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)70/70(100%)
Site4AgreementwithExpected result10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)10/10(100%)70/70(100%)
Total Agreement withExpectedresult40/40(100%)40/40(100%)40/40(100%)40/40(100%)40/40(100%)40/40(100%)40/40(100%)280/280(100%)
95% CI91.2 -100%91.2 -100%91.2 -100%91.2 -100%91.2 -100%91.2 -100%91.2 -100%98.7 -100%

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Limit of Detection:

Analytical Limit of Detection (LoD) of the L-DFA Reagent was addressed using dilution series of infected model cells. Model cells for parainfluenza virus types 1, 2, and 3 (ATCC strains C-35, Greer, and C243) and adenovirus (ATCC type 1) were diluted with non-infected cells to produce a suspension equivalent to 1,000 infected cells per milliliter. This level theoretically yields approximately 25 infected cells per 25-uL of suspension. This suspension was then serially diluted to a theoretical level of less than 1 cell per milliliter. (NOTE: This level was the target to begin with a low positive level. Actual starting levels vary, however, and are within 1 dilution of the 25 infected cell target level). 25-uL aliquots from each dilution level were spotted onto 10 replicate microscope slides, and then stained according to the instructions for use described in this product insert. Each cell spot was examined at 200X magnification. Results were reported as numbers of positive replicates for each set of 10. Analytical detection limits for each of the 4 analytes were defined as the lowest dilutions at which at least 9 out of 10 replicates were detected. LoD study results are summarized in Table 5.3 below:

TABLE 5.3:Limit of Detections of the D³ FastPoint L-DFA PIV/Adenovirus Reagent
Virus StrainInfected cells/mLNumber of replicates with positive cellsLOD determination
Adenovirus(ATCC type 1)100010/10100 infected cells/mL
20010/10
1009/10
505/10
251/10
12.50/10
60/10
30/10
1.50/10
0.80/10
hPIV-1(ATCC strain C-35)50010/10100 infected cells/mL
10010/10
506/10
252/10
12.51/10
60/10
30/10
1.50/10
0.80/10
0.40/10
hPIV-2(ATCC strain Greer)50010/1025 infected cells/mL
10010/10
5010/10
259/10
12.56/10
65/10
33/10
1.51/10
0.80/10

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D' FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit 10/30/2009 Section 05, Page 10 of 13

TABLE 5.3:Limit of Detections of the D³ FastPoint L-DFA PIV/Adenovirus Reagent
Virus StrainInfected cells/mLNumber of replicates with positive cellsLOD determination
hPIV-3(ATCC strain C243)100010/10
20010/10
10010/10
509/10
256/1050 infected cells/mL
12.52/10
60/10
30/10
1.50/10
0.80/10

Analytical reactivity (inclusivity):

Analytical reactivity (inclusivity) of the D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit was evaluated using 3 hPIV and 10 adenovirus strains. Low concentration infected cell suspensions (approximately 4% cells infected, 25 to 50 infected cells) were prepared for each viral strain. The suspensions were stained with the kit.

TABLE 5.4: Analytical Reactivity (inclusivity) of the D 3 FastPoint L-DFA PIV/Adenovirus Reagent onvarious hPIV and adenovirus strains
Parainfluenza and AdenovirusStrainsInfected Cell Concentration (asmultiples of the respectiveestablished LoD concentrationD3 FastPoint L-DFAPIV/Adenovirus Reagent Results
Parainfluenza 1 C-3510x LoD9 Golden-yellow fluorescent cells
Parainfluenza 2 Greer10x LoD11 Golden-yellow fluorescent cells
Parainfluenza 3 C-24310x LoD22 Golden-yellow fluorescent cells
Adenovirus 1 VR-110x LoD26 Apple-green fluorescent cells
Adenovirus 3 VR-310x LoD17 Apple-green fluorescent cells
Adenovirus 5 VR-510x LoD15 Apple-green fluorescent cells
Adenovirus 6 VR-610x LoD22 Apple-green fluorescent cells
Adenovirus 7 VR-710x LoD16 Apple-green fluorescent cells
Adenovirus 8 VR-136610x LoD29 Apple-green fluorescent cells
Adenovirus 10 VR-108710x LoD34 Apple-green fluorescent cells
Adenovirus VR-1410x LoD37 Apple-green fluorescent cells
Adenovirus Dewitt ATCC Strain10x LoD15 Apple-green fluorescent cells
Adenovirus 31 VR-110910x LoD42 Apple-green fluorescent cells

Clinical Performance:

Performance of the D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit testing direct respiratory specimens was established during prospective studies at 4 geographically diverse U.S. clinical laboratories during the 2009 respiratory virus seasons (January 2009 through March 2009). All specimens used in the studies meeting the inclusion and exclusion criteria represented excess, remnants of respiratory specimens that were prospectively collected from symptomatic

Sec05 FastPoint PIV-ADV 09OCT30

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individuals suspected of respiratory infection, and were submitted for routine care or analysis by each site, and that otherwise would have been discarded. Individual specimens were delinked from all patient identifiers and given a study sample code. All clinical sites were granted waivers of informed consent by their IRBs for this study.

Performance of the D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit was assessed and compared to a predetermined algorithm that used composite comparator methods. The composite comparator methods for parainfluenza virus and adenovirus consisted of Direct Specimen Fluorescent Antibody (DSFA) test with an FDA-cleared device and viral culture confirmation of all the negatives (as determined by the comparator DSFA test). "True" positive was defined as any sample that either tested positive by the comparator DSFA test or viral culture. "True" negative was defined as any sample that tested negative by both the comparator DSFA test and viral culture.

Prevalence of adenovirus and hPIV (human parainfluenza virus) within this population as determined by the D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit direct specimen testing is noted in Table 5.5 below:

TABLE 5.5: Parainfluenza Virus/Adenovirus Prevalence
AgeTotalSpecimensAdenovirus# positivehPIV# positive
Evaluated(prevalence)(prevalence)
0 to 1 month5501 (1.8%)
> 1 month to 2 years57711 (1.9%)29 (5.0%)
> 2 years to 12 years3911 (0.3%)6 (1.5%)
> 12 years to 21 years17302 (1.2%)
22 years to 30 years5701 (1.8%)
31 years to 40 years7100
41 years to 50 years5200
51 years to 60 years4600
61 years to 70 years3300
71 years to 80 years1600
81 years and above701 (14.3%)
Age Not Reported411 (2.4%)0
Total151913 (0.9%)40 (2.6%)

Tables 5.6 and 5.7 below show the study results of the NP wash/aspirate specimen type (Study Sites 1, 2, and 3 combined):

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TABLE 5.6: Parainfluenza Virus
Fresh nasal/nasopharyngealwash/aspirateComparator DSFA(negatives followed by culture with DFA)
DHI DSFAPositiveNegativeTotal
Positive23427
Negative2599601
Total25603628
95% CI
Sensitivity23/2592.0%74.0-99.0%
Specificity599/60399.3%98.3-99.8%
TABLE 5.7: Adenovirus
Fresh nasal/nasopharyngealwash/aspirateComparator DSFA(negatives followed by culture with DFA)
DHI DSFAPositiveNegativeTotal
Positive12012
Negative1619620
Total13619632
95% CI
Sensitivity12/1392.3%64.0-99.8%
Specificity619/619100%99.4-100%

Tables 5.8 and 5.9 below show the study results of the NP swab specimen type (Study Sites 3 and 4 combined):

TABLE 5.8: Parainfluenza Virus
Fresh nasal/nasopharyngealswabComparator DSFA(negatives followed by culture with DFA)
DHI DSFAPositiveNegativeTotal
Positive13013
Negative1668669
Total14668682
95% CI
Sensitivity13/1492.9%66.1-99.8%
Specificity668/668100%99.4-100%
TABLE 5.9: Adenovirus
Fresh nasal/nasopharyngealswabComparator DSFA(negatives followed by culture with DFA)
DHI DSFAPositiveNegativeTotal
Positive101
Negative0680680
Total1680681
95% CI
Sensitivity1/1100%N/A
Specificity680/680100%99.5-100%

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Note: The sensitivity performance of the D3 FastPoint PIV/ADV Kit detecting adenovirus from direct nasal/nasopharyngeal swab specimens has not been adequately established in the clinical study due to low adenovirus prevalence at the clinical study sites. However, the same MAb pool for adenovirus was validated in previous clinical trials for a number of FDA-cleared DSFA devices. Users may wish to further evaluate the sensitivity performance of this kit detecting adenovirus using prospective nasal/nasopharyngeal swab samples.

Overall at the four Study Sites, the performance results of the D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit, when compared to those of the comparator devices, D3 Ultra DFA Respiratory Virus Screening & ID Kit and D3 Duet DFA RSV/Respiratory Virus Screening Kit, demonstrate that the devices detect parainfluenza virus and adenovirus antigens in a similar manner.

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Image /page/13/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized depiction of an eagle or bird-like figure with three curved lines representing its wings or feathers. The logo is surrounded by the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" in a circular arrangement. The text is in all capital letters and is positioned around the upper half of the circle.

DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

Food and Drug Administration 10903 New Hampshire Avenue Document Mail Center-WO66-G609 Silver Spring, MD 20993-0002

Mr. Ronald H. Lollar Senior Director Product Realization, Management and Marketing Diagnostic Hybrids, Inc 1055 East State Street, Suite 100 Athens, Ohio 45701

DEC 2 3 2009

Re: K093415

Trade/Device Name: D3 FastPoint L- DFA Parainfluenza Virus/Adenovirus Identification Kit Regulation Number: 21 CFR & 866.3400 Regulation Name: Parainfluenza Virus Serological Reagents Regulatory Class: I Product Code: GQS, GNY Dated: October 30, 2009 Received: November 2, 2009

Dear Mr. Lollar:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21

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CFR Part 807); labeling (21 CFR Parts 801 and 809); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

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

If you desire specific information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5461. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 443-6597 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html

Sincerely yours,

Une Schuf for

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

Enclosure

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510(k) Number (if known): K093415

Device Name: D³ FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit

Indication for Use:

The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA Parainfluenza Virus/Adenovirus Identification Kit is intended for the qualitative identification of adenovirus and to screen for the presence of parainfluenza virus types 1, 2, and 3 in nasal and nasopharyngeal swabs and aspirates/washes specimens from patients with signs and symptoms of respiratory infection by direct detection of immunofluorescence using monoclonal antibodies (MAbs).

It is recommended that specimens found to be negative for parainfluenza virus and adenovirus after examination of the direct specimen result be confirmed by cell culture. Negative results do not preclude parainfluenza virus and adenovirus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.

Prescription Use X (Part 21 CFR 801 Subpart D)

AND/OR

Over-The-Counter Use (21 CFR 801 Subpart C)

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

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

Uve Schuf

Division Sign-Off

Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K093415

§ 866.3400 Parainfluenza virus serological reagents.

(a)
Identification. Parainfluenza virus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to parainfluenza virus in serum. The identification aids in the diagnosis of parainfluenza virus infections and provides epidemiological information on diseases caused by these viruses. Parainfluenza viruses cause a variety of respiratory illnesses ranging from the common cold to pneumonia.(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.