The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA Influenza A/Influenza B Virus Identification Kit is intended for the qualitative identification of influenza A virus and influenza B virus in nasal and nasopharyngeal swabs and aspirates/washes specimens from patients with signs and symptoms of respiratory infection by direct detection of immunofluorescence using monoclonal antibodies (MAbs).

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

Performance characteristics for influenza A virus detection and identification were established when influenza A (H3N2) and influenza A (H1N1) were the predominant influenza A strains circulating in the United States. Since influenza strains display antigenic drift and shift from year to year, performance characteristics may vary. If infection with a novel influenza A virus is suspected, based on clinical and epidemiological screening criteria communicated by public health authorities, collect specimens following appropriate infection control precautions and submit to state or local health departments, for testing. Viral culture should not be attempted in these cases unless a BSL 3+facility' is available to receive and culture specimens .

Device Description

The D3 FastPoint L-DFA Influenza A/Influenza B Virus Identification Kit (D3 FastPoint A/B Kit) uses a blend (called a "L-DFA Reagent") of viral antigen-specific murine monoclonal antibodies that are directly labeled with either R-PE (influenza A virus) or fluorescein (influenza B virus) for the rapid identification of influenza A virus and influenza B virus in nasal and nasopharyngeal swabs and aspirates/washes specimens from patients with signs and symptoms of respiratory infection.

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 re-suspension buffer and loaded onto a specimen slide well. The cells are examined using a fluorescence microscope. Cells infected with influenza A virus will exhibit goldenyellow fluorescence due to the PE. Cells infected with influenza B virus 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 a breakdown of the acceptance criteria and the study details for the D³ FastPoint L-DFA Influenza A/Influenza B Virus Identification Kit:

1. Table of Acceptance Criteria and Reported Device Performance:

The document doesn't explicitly state "acceptance criteria" in a quantitative format for clinical performance beyond the presented sensitivity and specificity. However, based on the clinical trial results, these would be the implied performance metrics.

Performance Metric	Target/Acceptance Criteria (Implied)	Reported Device Performance (Influenza A - Wash/Aspirate)	Reported Device Performance (Influenza A - Swab)	Reported Device Performance (Influenza B - Wash/Aspirate)	Reported Device Performance (Influenza B - Swab)
Clinical Sensitivity	High (e.g., >80%)	84.8% (95% CI: 73.9-92.5%)	87.7% (95% CI: 77.2-94.5%)	81.8% (95% CI: 48.2-97.7%)	87.9% (95% CI: 83.7-92.1%)
Clinical Specificity	High (e.g., >95%)	99.5% (95% CI: 98.5-99.9%)	99.8% (95% CI: 99.1-100%)	100.0% (95% CI: 99.4-100%)	99.8% (95% CI: 98.8-100%)

Analytical Performance (Reproducibility):

Performance Metric	Acceptance Criteria (Implied)	Reported Device Performance
Flu A detection	100% agreement expected	100% (120/120)
Flu B detection	100% agreement expected	100% (120/120)
Negative (no infected cells)	High agreement (e.g. >95%)	95% (38/40)
Total % Agreement	High (e.g. >95%)	99.3% (278/280)

Analytical Performance (Limit of Detection - LoD):

Virus Strain	Acceptance Criteria (Defined as lowest dilution with >= 9/10 replicates detected)	Reported Device Performance (LOD)
Influenza A (Victoria)	Detection in >= 9/10 replicates	50 infected cells/mL
Influenza B (Taiwan)	Detection in >= 9/10 replicates	50 infected cells/mL

Analytical Reactivity (Inclusivity):

The document states "MAbs are reactive with all listed strains" for both Influenza A and B. The acceptance criterion is implicitly 100% reactivity with the tested strains. The reported performance confirms this by showing detection of a specified number of fluorescent cells for all tested strains at 20x LoD.

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

Clinical Test Set Sample Size:
- Total Specimens Evaluated: 1519
- Nasal/Nasopharyngeal Wash/Aspirate (Influenza A): 637 specimens
- Nasal/Nasopharyngeal Wash/Aspirate (Influenza B): 628 specimens
- Nasal/Nasopharyngeal Swab (Influenza A): 690 specimens
- Nasal/Nasopharyngeal Swab (Influenza B): 711 specimens (Note: Summing these individual specimen counts for A and B across different sample types is not appropriate to get a single "total," as the same specimen might be tested for both A and B, and different subsets might be used for different sample types or sites.)
Data Provenance: Prospective collection from symptomatic individuals suspected of respiratory infection in 4 geographically diverse U.S. clinical laboratories during the 2009 respiratory virus season (January 2009 - March 2009). The specimens were "excess, remnants" that would have otherwise been discarded, implying real-world clinical samples.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts:

The document describes the ground truth as a "predetermined algorithm that used composite comparator methods" involving an FDA-cleared Direct Specimen Fluorescent Antibody (DSFA) test and viral culture confirmation. This method does not explicitly state the involvement of human experts establishing the ground truth beyond the likely requirement of trained laboratory personnel to perform and interpret the comparator tests. Therefore, details on the number and qualifications of experts for ground truth establishment are not provided in this document.

4. Adjudication Method for the Test Set:

The ground truth was established using a "composite comparator method":

"True" positive: Any sample that tested positive by either the comparator DSFA test OR viral culture.
"True" negative: Any sample that tested negative by BOTH the comparator DSFA test AND viral culture.
This is a form of adjudication by algorithm/composite reference standard rather than human consensus.

5. If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study was Done:

No, a Multi Reader Multi Case (MRMC) comparative effectiveness study of human readers with vs. without AI assistance was not done. The study compared the device (D3 FastPoint A/B Kit) directly against a composite reference standard, not against human readers. This device is a diagnostic kit, not an AI or imaging device that would typically involve human reader studies for interpretation.

6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) was Done:

Yes, the performance presented for the D3 FastPoint L-DFA Influenza A/Influenza B Virus Identification Kit is a standalone performance of the device (kit) itself, as read and interpreted by laboratory personnel according to its instructions. It is not an "algorithm" in the modern AI sense, but a lab kit where the reported accuracy reflects the kit's ability to correctly identify the viruses based on fluorescence.

7. The Type of Ground Truth Used:

The ground truth used was a composite reference standard consisting of:

An FDA-cleared Direct Specimen Fluorescent Antibody (DSFA) test.
Viral culture confirmation for all negatives from the comparator DSFA test.

8. The Sample Size for the Training Set:

The document does not explicitly mention a separate "training set" for the D3 FastPoint L-DFA Influenza A/Influenza B Virus Identification Kit in the context of machine learning or algorithm development. This device is a traditional in-vitro diagnostic kit. Its development would involve analytical studies and optimization (e.g., antibody selection, reagent formulation) rather than a separate "training set" in the AI sense.

However, the "Analytical Reactivity (inclusivity)" study, which tested the reactivity against 13 influenza A strains and 7 influenza B strains, could be considered part of the analytical development and validation process to ensure the kit's broad detection capabilities for known strains.

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

As there is no explicitly defined "training set" in the AI sense for this traditional diagnostic kit, the concept of establishing ground truth for a training set is not applicable here. The analytical studies (like inclusivity) involved preparing known infected cell suspensions with specified viral strains, where the "ground truth" was inherent in the preparation of these controlled samples.

Summary

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OCT 2 1 2009

D³ FastPoint L-DFA Influenza A/Influenza B Virus Identification Kit

9/14/2009 Page 1 of 14

Section 05, 510(k) Summary

Kog2882

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:

September 14, 2009

Device Name:

Trade name - D3 FastPoint L-DFA Influenza A/Influenza B Virus Identification Kit Common name - Influenza A/B virus DFA assay Classification name - Antisera, Cf, Influenza Virus A, B, C Product Code - GNW Regulation - 21 CFR 866.3330, Class I, Influenza virus serological reagents; Panel Microbiology (83)

Legally marketed devices to which equivalence is claimed:

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

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

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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 (Do Duet RSV Kit), is intended for the qualitative detection and identification of respiratory syncytial virus, while screening for influenza A virus, influenza B virus, adenovirus, and parainfluenza virus types 1, 2 and 3 viral antigens, in nasal and nasopharyngeal swabs and aspirates or in cell culture. The assay detects viral antigens by immunofluorescence using monoclonal antibodies (MAbs), from patients with signs and symptoms of respiratory infection.

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

Performance characteristics for influenza A virus detection and identification were established when influenza A (H3N2) and influenza A (H1N1) were the predominant influenza A strains circulating in the United States. Performance characteristics for influenza A virus detection and identification were established when influenza A H3N2 and influenza A H1N1 were the predominant influenza A strains circulating in the United States. When other influenza A viruses are emerging, performance characteristics may vary. If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to a state or local health department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

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Device Description:

Kit Components:

1. D³ FastPoint L-DFA Influenza A/Influenza B Reagent, 4.0-mL. One dropper bottle containing a mixture of PE-labeled murine monoclonal antibodies directed against influenza A virus antigens and FITC-labeled murine monoclonal antibodies directed against influenza B virus antigens. The buffered, stabilized, aqueous solution contains Evans Blue and propidium iodide as counter-stains and 0.1% sodium azide as preservative.
1. 40X PBS Concentrate, 25-mL. One bottle of 40X PBS concentrate containing 4% sodium azide (0.1% sodium azide after dilution to 1X using de-mineralized water).
1. Re-suspension Buffer, 6.0-mL. One bottle of a buffered glycerol solution and 0.1% sodium azide.
1. D3 FastPoint L-DFA Influenza A/Influenza B Antigen Control Slides, 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 influenza A virus, or influenza B virus. The negative wells contain non-infected cells. Each slide is intended to be stained only one time.

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Intended Use:

Characteristics	D³ FastPoint A/B Kit (Subject Device)	D³ Ultra Kit 510(k) #K061101	D³ Duet RSV Kit 510(k) # K081928
Intended Use	The Diagnostic Hybrids, Inc. device, D³ FastPoint L-DFA Influenza A/ Influenza B Virus Identification Kit is intended for the qualitative identification of influenza A virus and influenza B virus in nasal and	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.	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,
Characteristics	D³ FastPointA/B Kit(Subject Device)	D³ Ultra Kit510(k) #K061101	D³ Duet RSV Kit510(k) # K081928
	nasopharyngealswabs andaspirates/washesspecimens frompatients with signsand symptoms ofrespiratory infectionby direct detectionofimmunofluorescenceusing monoclonalantibodies (MAbs).It is recommended	respiratory syncytialvirus (RSV),adenovirus,parainfluenza 1,parainfluenza 2 andparainfluenza 3virus in respiratoryspecimens, by eitherdirect detection orcell culture method,byimmunofluorescenceusing monoclonalantibodies (MAbs).	adenovirus, andparainfluenza virustypes 1, 2 and 3 viralantigens, in nasal andnasopharyngeal swabsand aspirates or in cellculture. The assaydetects viral antigens byimmunofluorescenceusing monoclonalantibodies (MAbs),from patients with signsand symptoms ofrespiratory infection.
	that specimensfound to be negativefor influenza afterexamination of thedirect specimenresult be confirmedby cell culture.Negative results donot precludeinfluenza virusinfection and shouldnot be used as thesole basis fordiagnosis, treatmentor othermanagementdecisions.	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.	It is recommended thatspecimens found to benegative afterexamination of thedirect specimen resultbe confirmed by cellculture.Negative results do notpreclude influenza virusinfection and should notbe used as the sole basisfor diagnosis, treatmentor other managementdecisions.
Target Viruses	influenza A virus,influenza B virus	influenza A virus,influenza B virus,respiratorysyncytial virus,adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 3	influenza A virus,influenza B virus,respiratory syncytialvirus, adenovirus,parainfluenza virustype 1, parainfluenzavirus type 2,parainfluenza virustype 3
Monoclonal antibodies(MAbs)	The L-DFA Reagentcontains 4 MAbs toinfluenza A virus,influenza B virus	The RespiratoryVirus DFAScreening Reagentcontains 15 MAbs to	The RSV/RespiratoryVirus DFA ScreeningReagent contains 15MAbs to 7 different
Table 5.1: Characteristics of the D³ FastPoint L-DFA Influenza A/Influenza B Kit arecompared to those of the following Diagnostic Hybrids (DHI) predicate devices
Characteristics	D³ FastPointA/B Kit(Subject Device)	D³ Ultra Kit510(k) #K061101	D³ Duet RSV Kit510(k) # K081928
		respiratory viruses(influenza A virus,influenza B virus,respiratory syncytialvirus, adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 3)	(influenza A virus,influenza B virus,adenovirus,parainfluenza virus type1, parainfluenza virustype 2, parainfluenzavirus type 3), plus 2MAbs to respiratorysyncytial virus.
	Direct labeling,	Direct labeling,	Direct labeling,
Labeling method	- using R-Phycoerythrin (R-PE) to label theMAbs to influenzaA virus.		- using R-Phycoerythrin (R-PE)to label the MAbs torespiratory syncytialvirus.
	- using fluoresceinisothiocyanate(FITC) to labelinfluenza B virus,MAbs.	- using fluoresceinisothiocyanate(FITC) to label allMAbs withfluorescein.	- using fluoresceinisothiocyanate (FITC) tolabel all other MAbswith fluorescein.
R-Phycoerythrin-labeledMAbs	influenza A virus	None	respiratory syncytialvirus
Fluorescein-labeled MAbs	influenza B virus	influenza A virus,influenza B virus,respiratory syncytialvirus, adenovirus,parainfluenza virustype 1,parainfluenza virustype 2,parainfluenza virustype 3	influenza A virus,influenza B virus,adenovirus,parainfluenza virus type1, parainfluenza virustype 2, parainfluenzavirus type 3
Cell Fixative	Proprietary Non-Acetone basedsystem	Acetone	Acetone
Cell Counter-stain	Propidium Iodide,Evans Blue	Evans Blue	Evans Blue
Performance characteristics
Staining patterns	Influenza A and B:The fluorescence iscytoplasmic orbright nuclear orboth. Cells appear	Influenza A and B:The fluorescence iscytoplasmic, nuclearor both.Cytoplasmic	Influenza A and B:The fluorescence iscytoplasmic, nuclearor both. Cytoplasmicstaining is often
Characteristics of the D³ FastPoint L-DFA Influenza A/Influenza B Kit are compared to those of the following Diagnostic Hybrids (DHI) predicate devices
Characteristics	D³ FastPoint A/B Kit (Subject Device)	D³ Ultra Kit 510(k) #K061101	D³ Duet RSV Kit 510(k) # K081928
	round.Negative: Cells fluoresce red due to the Evans Blue counter-stain.Nuclei: Cell Nuclei fluoresce orange-red due to the Propidium Iodide counter-stain.	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 or both.Negative: Cells fluoresce red due to the Evans Blue counter-stain.	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 or both.Negative: Cells fluoresce red due to the Evans Blue counter-stain.
Analytical specificity (for influenza A virus strains; MAbs are reactive with all listed strains)	13 influenza A strains: Influenza A Mexico/4108/2009 (H1N1) from CDC, Influenza A California/07/2009 (H1N1) from CDC, Aichi (H3N2), Mal (H1N1), Hong Kong (H3N2), Denver (H1N1), Port Chalmers (H3N2), Victoria (H3N2), New Jersey (H1N1), WS (H1N1), PR (H1N1), Wisconsin (H3N2), WS (H1N1)	10 influenza A strains: Aichi (H3N2), Mal (H1N1), Hong Kong (H3N2), Denver (H1N1), Port Chalmers (H3N2), Victoria (H3N2), New Jersey (H1N1), WS (H1N1), PR,(H1N1), A/NWS/33 (H1N1)	10 influenza A strains:Aichi (H3N2), Mal (H1N1), Hong Kong (H3N2), Denver (H1N1), Port Chalmers (H3N2), Victoria (H3N2), New Jersey (H1N1), WS (H1N1), PR (H1N1), A/NWS/33 (H1N1)
Table 5.1: Characteristics of the D3 FastPoint L-DFA Influenza A/Influenza B Kit arecompared to those of the following Diagnostic Hybrids (DHI) predicate devices
Characteristics	D3 FastPointA/B Kit(Subject Device)A/NWS/33 (H1N1)	D3 Ultra Kit510(k) #K061101	D3 Duet RSV Kit510(k) # K081928
Analytical specificity (forInfluenza B virus strains;MAbs are reactive with alllisted strains)	7 influenza Bstrains: Hong Kong,Maryland, Mass,GL, Taiwan,B/Lee/40, Russia	7 influenza Bstrains: Hong Kong,Maryland, Mass,GL, Taiwan,B/Lee/40, Russia	7 influenza B strains:Hong Kong, Maryland,Mass, GL, Taiwan,B/Lee/40, Russia
Analyticalspecificity(cross-reactivitystudies; variousstrains ofmicroorganismsand cell lines)	Viruses	22	31	32
	Bacteria	22	18	25
	Chlamydia spp.	1	1	3
	Yeast	1	0	1
	Protozoan	1	0	1
	Cell lines	N/A	17	17

Technological Characteristics, Compared to Predicate Device:

www.cdc.gov

2 FDA Guidance Document: In Vitro Diagnostic Devices to Detect Influenza A Viruses: Labeling and Regulatory Path; Issued 4/10/2006

DHI-FastPoint Flu A-B_Scc05_510(k) Summary_09-0914

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D3 FastPoint L-DFA Respiratory Virus Identification Kit

Section 05, Page 5 of 14

DHI-FastPoint Flu A-B_Sec05_510(k) Summary_09-0914

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・

DHI-FastPoint Flu A-B_Scc05_510(k) Summary_09-0914

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D3 FastPoint L-DFA Respiratory Virus Identification Kit

・

9/14/2009 Section 05, Page 7 of 14

DHI-FastPoint Flu A-B_Sec05_510(k) Summary_09-0914

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D3 FastPoint L-DFA Respiratory Virus Identification Kit

Although the D FastPoint L-DFA Influenza A/Influenza B Reagent has been shown to detect the 2009 H1N1 virus in two culture isolates, the performance characteristics of this device with clinical specimens that are positive for the 2009 H1N1 influenza virus have not been established. The D3 FastPoint L-DFA Influenza A/Influenza B DFA Reagent can distinguish between influenza A and B viruses, but it cannot differentiate influenza subtypes.

Analytical 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 Influenza A/B panel consisted of the following:

a. Low level influenza A (Victoria strain) infected cells.
b. Low level influenza B (Taiwan strain) infected cells.
Low level influenza A (Victoria strain) infected cells mixed c. with mid level influenza B (Taiwan strain) infected cells.
d. Low level influenza B (Victoria strain) infected cells mixed with mid level influenza A (Victoria strain) infected cells.
e. Mid level non-infected (negative) cells.

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 x105 to 3.5 x105 total cells.

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

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

For the L-DFA Reagent, the combined data from the four Study Sites demonstrated reproducible detection of influenza A virus by the R-PE labeled MAbs and reproducible detection of influenza B virus by the FITClabeled MAbs. The presence of influenza A virus infected cells was reported in 100% (120/120) of the wells in which the infected cells were expected. The presence of influenza B virus infected cells was reported in 100% (120/120) of the wells in which the infected cells were expected. The absence of infected cells was reported in 95% (38/40) of the wells in which infected cells were not present. The total percent agreement for the L-DFA Reagent was 99.3% (278/280):

Table 5.2: Reproducibility Study Results using the L-DFA Reagent
Site	PanelMemberConcentration	NegativeNoinfectedcells	Flu ALowLevel4 to 10%infectedcells	Flu BLowLevel4 to 10%infectedcells	Mixed Infection		Mixed Infection		Total%Agreement
					Flu AMidLevel20 to30%infectedcells	Flu B LowLevel4 to 10%infectedcells	Flu ALow Level4 to 10%infectedcells	Flu B MidLevel20 to 30%infectedcells
Site1	AgreementwithExpected result	8/10(80%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	68/70(97.1%)
Site2	AgreementwithExpected result	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	70/70(100%)
Site3	AgreementwithExpected result	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	70/70(100%)
Site4	AgreementwithExpected result	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	10/10(100%)	70/70(100%)
	Total Agreement withExpectedresult	38/40(95%)	40/40(100%)	40/40(100%)	40/40(100%)	40/40(100%)	40/40(100%)	40/40(100%)	278/280(99.3%)
	95% CI	83.1 -99.4%	91.2 -100%	91.2 -100%	91.2 -100%	91.2 -100%	91.2 -100%	91.2 -100%	97.4 -99.9%

Limit of Detection

Analytical Limit of Detections (LoDs) of the L-DFA Reagent was addressed using dilution series of infected model cells. Model cells for influenza A virus

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(ATCC Victoria strain), influenza B virus (ATCC Taiwan strain) were diluted with non-infected cells to produce a suspension equivalent to 1,000 infected cells per milliliter. This level theoretically vields 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-μL aliquots from each dilution level were spotted onto 10 replicate microscope slides, and then stained according to the instructions for use described in this product insert. Each cell 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 8 analytes were defined as the lowest dilutions at which at least 9 out of 10 replicates were detected. LoD study results are summarized in Table 5.3 below:

Table 5.3: Limit of Detections of the L-DFA Reagent
Virus Strain	Infected cells/mL	Number of replicates with positive cells	LOD determination
Influenza A(ATCC Victoriastrain)	500	10/10	50 infected cells/mL
	100	10/10
	50	10/10
	25	5/10
	12.5	3/10
	6	2/10
	3	0/10
	1.5	2/10
	0.8	0/10
	0.4	0/10
Influenza B(ATCC Taiwan strain)	2000	10/10	50 infected cells/mL
	400	10/10
	200	10/10
	100	10/10
	50	10/10
	25	7/10
	12.5	4/10
	6	2/10
	3	0/10
	1.5	0/10

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Analytical reactivity (inclusivity)

Analytical reactivity (inclusivity) of the L-DFA Reagent was evaluated using 13 influenza A virus and 7 influenza B virus strains. Low concentration infected cell suspensions (approximately 4% cells infected, 25-50 infected cells) were prepared for each viral strain. The suspensions were stained with the L-DFA Reagent.

Table 5.4: Analytical Reactivity (inclusivity) of the L-DFA Reagent on various influenza A virus and influenzaB virus strains
Influenza Strains	Infected Cell Concentration(as multiples of the respectiveestablished LoD concentration	L-DFA Reagent Results
Influenza A Mexico/4108/2009(H1N1) from CDC*	20x LoD	19 Golden-yellow fluorescent cells
Influenza A California/07/2009(H1N1) from CDC*	20x LoD	26 Golden-yellow fluorescent cells
Influenza A Wisconsin/56/2005(H3N2)	20x LoD	39 Golden-yellow fluorescent cells
Influenza A WS, VR-1520 (H1N1)	20x LoD	67 Golden-yellow fluorescent cells
Influenza A Hong Kong, VR-544(H3N2)	20x LoD	13 Golden-yellow fluorescent cells
Influenza A New Jersey, VR-897(H1N1)	20x LoD	15 Golden-yellow fluorescent cells
Influenza A A/NWS/33 (H1N1)	20x LoD	10 Golden-yellow fluorescent cells
Influenza A Victoria, VR-822 (H3N2)	20x LoD	10 Golden-yellow fluorescent cells
Influenza A PR, VR-95 (H1N1)	20x LoD	20 Golden-yellow fluorescent cells
Influenza A Port Chalmers, VR-810(H3N2)	20x LoD	8 Golden-yellow fluorescent cells
Influenza A Aichi, VR-547 (H3N2)	20x LoD	28 Golden-yellow fluorescent cells
Influenza A Denver, VR-546 (H1N1)	20x LoD	30 Golden-yellow fluorescent cells
Influenza A Mal, VR-98 (H1N1)	20x LoD	21 Golden-yellow fluorescent cells
Influenza B GL/1739/54, VR-103	20x LoD	13 Apple-green fluorescent cells
Influenza B Taiwan/2/62, VR-295	20x LoD	44 Apple-green fluorescent cells
Influenza B Hong Kong/5/72, VR-823	20x LoD	21 Apple-green fluorescent cells
Influenza B Maryland/1/59, VR-296	20x LoD	22 Apple-green fluorescent cells
Influenza B Russia, VR-790	20x LoD	36 Apple-green fluorescent cells
Influenza B B/Lee/40	20x LoD	41 Apple-green fluorescent cells
Influenza B Massachusetts, VR-523	20x LoD	67 Apple-green fluorescent cells

Although the D3 FastPoint L-DFA Influenza A/Influenza B Reagent has been shown to detect the 2009 H1N1 influenza virus in two culture isolates, the performance characteristics of this device with clinical specimens that are positive for the 2009 H1N1 influenza virus have not been established. The D3 FastPoint L-DFA Influenza A/Influenza B DFA Reagent can distinguish between influenza A and B viruses, but it cannot differentiate influenza subtypes.

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Clinical Performance:

Performance of the D FastPoint A/B Kit testing direct respiratory specimens were established during prospective studies at 4 geographically diverse U.S. clinical laboratories during the 2009 respiratory virus seasons (January 2009 - March 2009). All specimens used in the studies meeting the inclusion and exclusion criteria represented excess, remnants of respiratory specimens that were prospectively collected from symptomatic individuals suspected of respiratory infection, and were submitted for routine care or analysis by each site, and that otherwise would have been discarded. Individual specimens were delinked from all patient identifiers and given a study sample code. All clinical sites were granted waivers of informed consent by their IRBs for this study.

Performance of the D3 FastPoint A/B Kit was assessed and compared to a predetermined algorithm that used composite comparator methods. The composite comparator methods for 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 Influenza A/B viruses within this population as determined by the D3 FastPoint A/B Kit direct specimen testing is noted in Table 5.5 below:

Table 5.5: Influenza A/B Virus Prevalence
Age	TotalSpecimensEvaluated	Flu A# positive(prevalence)	Flu B# positive(prevalence)
0 - 1 month	55	0	0
> 1 month to 2 years	577	27 (4.7%)	20 (3.5%)
> 2 years to 12 years	391	43 (11.0%)	104 (26.6%)
> 12 years to 21 years	173	19 (11.0%)	41 (23.7%)
22 years to 30 years	57	3 (5.3%)	14 (24.6%)
31 years to 40 years	71	9 (12.7%)	9 (12.7%)
41 years to 50 years	52	5 (9.6%)	5 (9.6%)
51 years to 60 years	46	3 (6.5%)	3 (6.5%)
61 years to 70 years	33	2 (6.1%)	2 (6.1%)
71 years to 80 years	16	2 (12.5%)	1 (6.3%)
81 years and above	7	0	0
Age Not Reported	41	2 (4.9%)	14 (34.1%)
Total	1519	115 (7.6%)	213 (14.0%)

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

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: 1000

D3 FastPoint L-DFA Respiratory Virus Identification Kit

9/14/2009 Section 05, Page 13 of 14

Table 5.6: Influenza A
Fresh nasal/nasopharyngealwash/aspirate	Comparator DSFA(negatives followed by culture with DFA)
DHI DSFA	Positive	Negative	Total
Positive	56	3	59
Negative	10	568	578
Total	66	571	637
			95% CI
Sensitivity	56/66	84.8%	73.9-92.5%
Specificity	568/571	99.5%	98.5-99.9%

Table 5.7: Influenza B
Fresh nasal/nasopharyngealwash/aspirate	Comparator DSFA(negatives followed by culture with DFA)
DHI DSFA	Positive	Negative	Total
Positive	9	0	9
Negative	2	617	619
Total	11	617	628
			95% CI
Sensitivity	9/11	81.8%	48.2-97.7%
Specificity	617/617	100.0%	99.4-100%

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

Table 5.8: Influenza A
Fresh nasal/nasopharyngealswab	Comparator DSFA(negatives followed by culture with DFA)
DHI DSFA	Positive	Negative	Total
Positive	57	1	58
Negative	8	624	632
Total	65	625	690
			95% CI
Sensitivity	57/65	87.7%	77.2-94.5%
Specificity	624/625	99.8%	99.1-100%

Table 5.9: Influenza B
Fresh nasal/nasopharyngealswab	Comparator DSFA(negatives followed by culture with DFA)
DHI DSFA	Positive	Negative	Total
Positive	203	1	204
Negative	28	479	507
Total	231	480	711
			95% CI
Sensitivity	203/231	87.9%	83.7-92.1%
Specificity	479/480	99.8%	98.8-100%

DHI-FastPoint Flu A-B_Sec05_510(k) Summary_09-0914

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D3 FastPoint L-DFA Respiratory Virus Identification Kit

Section 05, Page 14 of 14

Overall at the four Study Sites, the performance results of the D3 FastPoint L-DFA Influenza A/Influenza B Virus Identification Kit, when compared to those of the comparator devices, D3 Ultra Kit, and D3 Duet RSV Kit, demonstrate that the devices detect influenza A virus and influenza B virus antigens in a similar manner.

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Image /page/14/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized depiction of an eagle or bird-like figure with three curved lines representing its body and wings. The logo is encircled by the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" in a circular arrangement.

DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

Food and Drug Administration 10903 New Hampshire Avenue Building 66 Silver Spring, MD 20993 ・

OCT 2 1 2009

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

Re: K092882

Trade/Device Name: D3 FastPoint L-DFA Influenza A/ Influenza B Virus Identification Kit

Regulation Number: 21 CFR 866.3330 Regulation Name: Influenza virus serological reagents Regulatory Class: Class I Product Code: GNX Dated: September 14, 2009 Received: September 18, 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

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

If you desire specific information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 594-3084. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 443-6597 or at its Internet address http://www.fda.gov/cdrh/dsma/dsmamain.html.

Sincerely yours,

Siyastgum

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): K092882

Device Name: D2 FastPoint L-DFA Influenza A/Influenza B Virus Identification Kit

Indications For Use:

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

Performance characteristics for influenza A virus detection and identification were established when influenza A (H3N2) and influenza A (H1N1) were the predominant influenza A strains circulating in the United States. Since influenza strains display antigenic drift and shift from year to year, performance characteristics may vary. If infection with a novel influenza A virus is suspected, based on clinical and epidemiological screening criteria communicated by public health authorities, collect specimens following appropriate infection control precautions and submit to state or local health departments, for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens .

AND/OR Over-The-Counter Use Prescription Use X (21 CFR 807 Subpart C) (Part 21 CFR 801 Subpart D)

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

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

signature
Division Sign-Off

Office of In Vitro Diagnostic Device Evaluation and Safety
Page 1 of 1

510(k)	k09 2882
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1<www.cdc.gov>
2FDA Guidance Document: In Vitro Diagnostic Devices to Detect Influenza A Viruses: Labeling and Regulatory Path; Issued 4/10/2006

Regulation Number and Section

§ 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.