Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description and intended use focus on immunofluorescence using monoclonal antibodies and fluorescence microscopy for direct detection. There is no mention of any computational analysis, algorithms, or learning processes that would indicate the use of AI or ML. The performance evaluation is based on traditional metrics like sensitivity and specificity compared to comparator methods.

Therapeutic

No
The device is intended for the qualitative identification of various respiratory viruses, serving as a diagnostic tool rather than providing direct therapy or treatment.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the device "is intended for the qualitative identification of influenza A virus, influenza B virus, respiratory syncytial virus, human metapneumovirus, adenovirus and to screen for the presence of parainfluenza virus types 1, 2, and 3". This aligns with the definition of a diagnostic device, which is used to identify the presence of diseases or conditions.

SaMD (Software as a Medical Device)

No

The device is a kit containing physical reagents (monoclonal antibodies, buffers, stains) used for immunofluorescence staining of respiratory specimens. The analysis is performed using a fluorescence microscope, which is a hardware component. The description clearly outlines the physical components and the manual laboratory procedure.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The intended use explicitly states that the device is for the "qualitative identification of influenza A virus, influenza B virus, respiratory syncytial virus, human metapneumovirus, 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". This is a clear diagnostic purpose, identifying specific pathogens in human specimens.
Specimen Type: The device is designed to be used with human specimens (nasal and nasopharyngeal swabs and aspirates/washes).
Methodology: The device uses immunofluorescence with monoclonal antibodies to detect viral antigens directly in the specimen. This is a common in vitro diagnostic technique.
Purpose: The results are intended to aid in the diagnosis of respiratory virus infections in symptomatic patients. While negative results require confirmation and should not be the sole basis for diagnosis, the primary purpose is diagnostic identification.

The description clearly aligns with the definition of an In Vitro Diagnostic device, which is a medical device intended to be used in vitro for the examination of specimens derived from the human body solely or principally for the purpose of providing information concerning a physiological state, state of health or disease, or congenital abnormality.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA Respiratory Virus Identification Kit is intended for the qualitative identification of influenza A virus, influenza B virus, respiratory syncytial virus, human metapneumovirus, 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 influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus or parainfluenza viruses after examination of the direct specimen result be confirmed by cell culture. Specimens found to be negative for human metapneumovirus after examination of the direct specimen results should be confirmed by an FDA cleared human metapneumovirus molecular assay. 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 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.

Product codes

OMG, LKT, GNX, GOS, GNY

Device Description

The D3 FastPoint L-DFA Respiratory Virus Identification Kit uses three blends (each called a "L-DFA Reagent") of viral antigen-specific murine monoclonal antibodies that are directly labeled with either R-PE (influenza A virus, respiratory syncytial virus, and parainfluenza virus) or fluorescein (influenza B virus, metapneumovirus, and adenovirus) for the rapid identification of respiratory viruses in nasal and nasopharyngeal swabs and aspirates from patients with signs and symptoms of respiratory infection.

Kit Components:

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.
D3 FastPoint L-DFA RSV/MPV Reagent, 4.0-mL. One dropper bottle containing a mixture of PE-labeled murine monoclonal antibodies directed against respiratory syncytial virus antigens and FITC-labeled murine monoclonal antibodies directed against metapneumovirus antigens. The buffered, stabilized, aqueous solution contains Evans Blue and propidium iodide as counter-stains and 0.1% sodium azide as preservative.
D3 FastPoint L-DFA PIV/Adenovirus Reagent, 4.0-mL. One dropper bottle containing a mixture of PE-labeled murine monoclonal antibodies directed against parainfluenza virus types 1, 2, or 3 antigens and FITClabeled 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 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 0.1% sodium azide.
D3 FastPoint L-DFA Respiratory Virus Antigen Control Slides, 5-slides. Five individually packaged control slides containing 6 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 virus, influenza B virus, respiratory syncytial virus, metapneumovirus, parainfluenza virus, and adenovirus. The negative wells contain non-infected cells. Each slide is intended to be stained only one time.

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 in 3 separate vials, each containing one of the 3 above reagents. 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 influenza A virus, respiratory syncytial virus, or parainfluenza virus types 1, 2 and 3 will exhibit golden-yellow fluorescence due to the PE. Cells infected with influenza B virus, metapnemovirus or 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.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

nasal and nasopharyngeal swabs and aspirates/washes specimens

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Not Found

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Not Found

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Analytical Performance:
Precision/Reproducibility:
Assay precision, intra-assay variability and inter assay variability were assessed with 3 panels of proficiency-level antigen control slides. Each of the 3 reproducibility panels consisted of 5 randomized panel members. Each panel was tested daily in two separate runs for 5-days by four different laboratories (40 total runs).
For the D3 FastPoint L-DFA Influenza A/Influenza B Reagent, the total percent agreement was 99.3% (278/280).
For the D3 FastPoint L-DFA RSV/hMPV Reagent, the total percent agreement was 100% (280/280).
For the D3 FastPoint L-DFA HPIV/Adenovirus Reagent, the total percent agreement was 100% (280/280).

Limit of Detection:
Analytical Limit of Detections (LoDs) of the D3 FastPoint L-DFA Reagents was addressed using dilution series of infected model cells. Model cells for 8 characterized respiratory virus isolates were diluted with non-infected cells to produce a suspension equivalent to 1,000 infected cells per milliliter. This suspension was then serially diluted. 25-μL aliquots from each dilution level were spotted onto 10 replicate microscope slides, and then stained. Each cell snot was examined at 200x magnification. 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.
Flu A (ATCC Victoria strain): 50 infected cells/mL
Flu B (ATCC Taiwan strain): 50 infected cells/mL
RSV (ATCC Washington strain): 100 infected cells/mL
hMPV A1 (Clinical strain): 100 infected cells/mL
Adenovirus (ATCC type 1): 100 infected cells/mL
HPIV-1 (ATCC strain C-35): 100 infected cells/mL
HPIV-2 (ATCC strain Greer): 25 infected cells/mL
HPIV-3 (ATCC strain C243): 50 infected cells/mL

Analytical reactivity (inclusivity):
Evaluated for D3FastPoint L-DFA Influenza A/Influenza B Reagent 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 and stained. All tested strains showed positive reactivity with golden-yellow or apple-green fluorescent cells ranging from 8 to 67.
Evaluated for D3 FastPoint L-DFA RSV/hMPV DFA Reagent using 3 RSV virus and 4 hMPV virus strains. Low concentration infected cell suspensions (approximately 4% cells infected, 25-50 infected cells) were prepared for each viral strain and stained. All tested strains showed positive reactivity with golden-yellow or apple-green fluorescent cells ranging from 22 to 37.
Evaluated for D3 FastPoint L-DFA HPIV/Adenovirus DFA Reagent using 3 HPIV virus and 10 Adenovirus strains. Low concentration infected cell suspensions (approximately 4% cells infected, 25-50 infected cells) were prepared for each viral strain and stained. All tested strains showed positive reactivity with golden-yellow or apple-green fluorescent cells ranging from 9 to 42.

Clinical Performance:
Prospective studies at 4 geographically diverse U.S. clinical laboratories during the 2009 respiratory virus seasons (January 2009 -- March 2009). Specimens were excess, remnants of respiratory specimens from symptomatic individuals suspected of respiratory infection.
Comparison was made to a predetermined algorithm that used composite comparator methods: Direct Specimen Fluorescent Antibody (DSFA) test with an FDA cleared device and viral culture confirmation of all negatives for influenza A, influenza B, RSV, parainfluenza, and adenovirus. For hMPV, the composite comparator included DSFA with an FDA cleared device, and confirmation of all negative specimens using a validated hMPV real-time RT-PCR followed by bidirectional sequencing analysis comparator assay.

Summary of results for NP wash/aspirate specimen type (Sites 1, 2, and 3 combined):
Flu A: Sensitivity 84.8% (56/66), Specificity 99.5% (568/571)
Flu B: Sensitivity 81.8% (9/11), Specificity 100.0% (617/617)
RSV: Sensitivity 98.6% (204/207), Specificity 99.8% (462/463)
Adenovirus: Sensitivity 92.3% (12/13), Specificity 100.0% (619/619)
HPIV: Sensitivity 92.0% (23/25), Specificity 99.3% (599/603)
hMPV: Sensitivity 68.8% (55/80), Specificity 100.0% (614/614)

Summary of results for NP swab specimen type (Sites 3 and 4 combined):
Flu A: Sensitivity 87.7% (57/65), Specificity 99.8% (623/624)
Flu B: Sensitivity 87.9% (203/231), Specificity 99.8% (478/479)
RSV: Sensitivity 97.5% (39/40), Specificity 100.0% (646/646)
Adenovirus: Sensitivity 100.0% (1/1), Specificity 100.0% (679/679). (Note: low adenovirus prevalence at sites, further evaluation recommended)
HPIV: Sensitivity 92.9% (13/14), Specificity 100.0% (667/667)
hMPV: Sensitivity 54.5% (24/44), Specificity 100.0% (631/631)

Overall, the performance results of the D3 FastPoint L-DFA Respiratory Virus Identification Kit, when compared to those of the comparator devices, demonstrate that the devices detect respiratory virus antigens in a similar manner.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

NP wash/aspirate specimen type:
Flu A: Sensitivity 84.8% (56/66), Specificity 99.5% (568/571)
Flu B: Sensitivity 81.8% (9/11), Specificity 100.0% (617/617)
RSV: Sensitivity 98.6% (204/207), Specificity 99.8% (462/463)
Adenovirus: Sensitivity 92.3% (12/13), Specificity 100.0% (619/619)
HPIV: Sensitivity 92.0% (23/25), Specificity 99.3% (599/603)
hMPV: Sensitivity 68.8% (55/80), Specificity 100.0% (614/614)

NP swab specimen type:
Flu A: Sensitivity 87.7% (57/65), Specificity 99.8% (623/624)
Flu B: Sensitivity 87.9% (203/231), Specificity 99.8% (478/479)
RSV: Sensitivity 97.5% (39/40), Specificity 100.0% (646/646)
Adenovirus: Sensitivity 100.0% (1/1), Specificity 100.0% (679/679)
HPIV: Sensitivity 92.9% (13/14), Specificity 100.0% (667/667)
hMPV: Sensitivity 54.5% (24/44), Specificity 100.0% (631/631)

Predicate Device(s)

D3 Ultra DFA Respiratory Virus Screening & ID Kit (K061101), D3 Duet DFA RSV/Respiratory Virus Screening Kit (K081928), D3 DFA Metapneumovirus Identification Kit (K090073)

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.

(a)
Identification. A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B;
(2) Influenza A subtype H1 and Influenza A subtype H3;
(3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B;
(4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus;
(5) Human Metapneumovirus;
(6) Rhinovirus; and
(7) Adenovirus.
(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;”
(2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and
(3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.

Summary

0

6691171

D3 FastPoint L-DFA Respiratory Virus Identification Kit

8/24/2009 Page 1 of 23

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:

April 17, 2009

Device Name:

Trade name - D3 FastPoint L-DFA Respiratory Virus Identification Kit Common name - Respiratory 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 after examination of the direct specimen

1

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)

The Diagnostic Hybrids, Inc. device, D3 Duet DFA RSV/Respiratory Virus Screening Kit (D3 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

2

attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

D3 DFA Metapneumovirus Identification Kit (K090073)

The Diagnostic Hybrids, Inc. device, D3 DFA Metapneumovirus Identification Kit (DJ MPV Kit), is intended for the qualitative detection and identification of human metapneumovirus (hMPV) in nasal and nasopharyngeal swabs and aspirates/washes or cell culture. The assay detects hMPV antigens by immunofluorescence using a blend of three monoclonal antibodies (MAbs), from patients with signs and symptoms of acute respiratory infection. This assay detects but is not intended to differentiate the four recognized genetic sub-lineages of hMPV.

Negative results do not preclude hMPV infection and should not be used as the sole basis for diagnosis, treatment or other management decisions. It is recommended that specimens found to be negative after examination of the direct specimen results be confirmed by an FDAcleared hMPV molecular assay.

Device Description:

The D3 FastPoint L-DFA Respiratory Virus Identification Kit uses three blends (each called a "L-DFA Reagent") of viral antigen-specific murine monoclonal antibodies that are directly labeled with either R-PE (influenza A virus, respiratory syncytial virus, and parainfluenza virus) or fluorescein (influenza B virus, metapneumovirus, and adenovirus) for the rapid identification of respiratory viruses in nasal and nasopharyngeal swabs and aspirates from patients with signs and symptoms of respiratory infection.

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. D3 FastPoint L-DFA RSV/MPV Reagent, 4.0-mL. One dropper bottle containing a mixture of PE-labeled murine monoclonal antibodies directed against respiratory syncytial virus antigens and FITC-labeled murine monoclonal antibodies directed against metapneumovirus antigens. The buffered, stabilized, aqueous solution contains Evans Blue and propidium iodide as counter-stains and 0.1% sodium azide as preservative.

3

1. D3 FastPoint L-DFA PIV/Adenovirus Reagent, 4.0-mL. One dropper bottle containing a mixture of PE-labeled murine monoclonal antibodies directed against parainfluenza virus types 1, 2, or 3 antigens and FITClabeled 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.
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 Respiratory Virus Antigen Control Slides, 5-slides. Five individually packaged control slides containing 6 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 virus, influenza B virus, respiratory syncytial virus, metapneumovirus, parainfluenza virus, and adenovirus. The negative wells contain noninfected cells. Each slide is intended to be stained only one time.

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 in 3 separate vials, each containing one of the 3 above reagents. 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 influenza A virus, respiratory syncytial virus, or parainfluenza virus types 1, 2 and 3 will exhibit goldenyellow fluorescence due to the PE. Cells infected with influenza B virus, metapnemovirus or 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.

4

Intended Use:

The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA Respiratory Virus Identification Kit is intended for the qualitative identification of influenza A virus, influenza B virus, respiratory syncytial virus, human metapneumovirus, 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 influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus or parainfluenza viruses after examination of the direct specimen result be confirmed by cell culture. Specimens found to be negative for human metapneumovirus after examination of the direct specimen results should be confirmed by an FDA cleared human metapneumovirus molecular assay. 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 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.2

Technological Characteristics, Compared to Predicate Device:

| Table 5.1: Characteristics of the D³ FastPoint L-DFA Kit are compared to those of the following

Diagnostic Hybrids (DHI) predicate devices
Characteristics	D³ FastPoint L-DFA Kit Subject Device	D³ Ultra Kit
510(k) #K061101	D³ Duet RSV Kit
510(k) # K081928	D³ MPV Kit
510(k) # K090073
Intended Use	The Diagnostic
Hybrids, Inc. device,
D³ FastPoint L-DFA
Respiratory Virus
Identification Kit is
intended for the	The Diagnostic
Hybrids, Inc. D³
Ultra ™ DFA (direct
fluorescent
antibody)
Respiratory Virus	The Diagnostic
Hybrids, Inc. device,
D³ Duet DFA
RSV/Respiratory
Virus Screening Kit,
is intended for the	The Diagnostic
Hybrids, Inc. device
D³ DFA
Metapneumovirus
Identification Kit, is
intended for the
Characteristics	D³ FastPoint L-DFA Kit Subject Device	D³ Ultra Kit 510(k) #K061101	D³ Duet RSV Kit 510(k) # K081928	D³ MPV Kit 510(k) # K090073
	qualitative
identification of
influenza A
virus, influenza B
virus, respiratory
syncytial virus,
human
metapneumovirus,
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 influenza A
virus, influenza B
virus, respiratory
syncytial virus,
adenovirus or
parainfluenza
viruses after
examination of the
direct specimen
result be confirmed
by cell culture.

Specimens found to
be negative for
human
metapneumovirus
after examination of
the direct specimen
results should be
confirmed by an
FDA cleared human | 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 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. | 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. | qualitative detection
and identification of
human
metapneumovirus
(hMPV) in nasal and
nasopharyngeal
swabs and
aspirates/washes or
cell culture. The
assay detects hMPV
antigens by
immunofluorescenc
e using a blend of
three monoclonal
antibodies (MAbs),
from patients with
signs and symptoms
of acute respiratory
infection. This
assay detects but is
not intended to
differentiate the fou
recognized genetic
sub-lineages of
hMPV.

Negative results do
not preclude hMPV
infection and should
not be used as the
sole basis for
diagnosis, treatment
or other
management
decisions. It is
recommended that
specimens found to
be negative after
examination of the
direct specimen
results be confirmed
by an FDA-cleared
hMPV molecular
assay. |
| Table 5.1: Characteristics of the D³ FastPoint L-DFA Kit are compared to those of the following
Diagnostic Hybrids (DHI) predicate devices | | | | |
| Characteristics | D³ FastPoint L-DFA
Kit Subject Device | D³ Ultra Kit
510(k) #K061101 | D³ Duet RSV Kit
510(k) # K081928 | D³ MPV Kit
510(k) # K090073 |
| | metapneumovirus
molecular assay.
Negative results do
not preclude
respiratory virus
infection and should
not be used as the
sole basis for
diagnosis, treatment
or other
management
decisions. | | | |
| Target Viruses | influenza A virus,
influenza B virus,
respiratory syncytial
virus,
metapneumovirus,
adenovirus,
parainfluenza virus
type 1,
parainfluenza virus
type 2,
parainfluenza virus
type 3 | influenza A virus,
influenza B virus,
respiratory
syncytial virus,
adenovirus,
parainfluenza virus
type 1,
parainfluenza virus
type 2,
parainfluenza virus
type 3 | influenza A virus,
influenza B virus,
respiratory
syncytial virus,
adenovirus,
parainfluenza virus
type 1,
parainfluenza virus
type 2,
parainfluenza virus
type 3 | metapneumovirus |
| Monoclonal antibodies
(MAbs) | The D³ FastPoint L-
DFA Reagents
contain 18 MAbs to
8 different
respiratory viruses
(influenza A virus,
influenza B virus,
respiratory syncytial
virus,
metapneumovirus,
adenovirus,
parainfluenza virus
type 1,
parainfluenza virus
type 2,
parainfluenza virus
type 3) | The Respiratory
Virus DFA
Screening Reagent
contains 15 MAbs to
7 different
respiratory viruses
(influenza A virus,
influenza B virus,
respiratory syncytial
virus, adenovirus,
parainfluenza virus
type 1,
parainfluenza virus
type 2,
parainfluenza virus
type 3) | The
RSV/Respiratory
Virus DFA
Screening Reagent
contains 15 MAbs to
7 different
respiratory viruses
(influenza A virus,
influenza B virus,
adenovirus,
parainfluenza virus
type 1, parainfluenza
virus type 2,
parainfluenza virus
type 3), plus 2 MAbs
to respiratory
syncytial virus. | The
Metapneumovirus
DFA Reagent
contains 3 MAbs to
metapneumovirus |
| Labeling method | Direct labeling,

using R-
Phycoerythrin (R-
PE) to label the | Direct labeling, | Direct labeling,
using R-
Phycoerythrin (R-
PE) to label the | Direct labeling, |
| Table 5.1: Characteristics of the D³ FastPoint L-DFA Kit are compared to those of the following
Diagnostic Hybrids (DHI) predicate devices. | | | | |
| Characteristics | D³ FastPoint L-DFA
Kit Subject Device | D³ Ultra Kit
510(k) #K061101 | D³ Duet RSV Kit
510(k) # K081928 | D³ MPV Kit
510(k) # K090073 |
| | A virus, RSV and
parainfluenza virus
types 1, 2 and 3. | | syncytial virus. | |
| | - using fluorescein
isothiocyanate
(FITC) to label
influenza B virus,
metapneumovirus
and adenovirus
MAbs with
fluorescein. | - using fluorescein
isothiocyanate
(FITC) to label all
MAbs with
fluorescein. | - using fluorescein
isothiocyanate
(FITC) to label all
other MAbs with
fluorescein. | - using fluorescein
isothiocyanate
(FITC) to label all
MAbs with
fluorescein. |
| R-Phycoerythrin-labeled
MAbs | influenza A virus,
respiratory syncytial
virus, parainfluenza
virus type 1,
parainfluenza virus
type 2,
parainfluenza virus
type 3 | None | respiratory syncytial
virus | None |
| Fluorescein-labeled MAbs | influenza B virus,
metapneumovirus,
adenovirus | influenza A virus,
influenza B virus,
respiratory syncytial
virus, adenovirus,
parainfluenza virus
type 1,
parainfluenza virus
type 2,
parainfluenza virus
type 3 | influenza A virus,
influenza B virus,
adenovirus,
parainfluenza virus
type 1, parainfluenza
virus type 2,
parainfluenza virus
type 3 | metapneumovirus |
| Cell Fixative | Proprietary Non-
Acetone based
system | Acetone | Acetone | Acetone |
| Cell Counter-stain | Propidium Iodide,
Evans Blue | Evans Blue | Evans Blue | Evans Blue |
| Performance characteristics | | | | |
| Staining patterns | Influenza A and B:
The fluorescence is
cytoplasmic or
bright nuclear or
both. Cells appear
round.
Respiratory
Syncytial Virus:
The fluorescence is
cytoplasmic. Cells
appear round. | 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 | 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 | Metapneumovirus:
The fluorescence is
cytoplasmic and
punctate with small
inclusions in the
syncytia.
Negative: Entire
cell fluoresce red
due to the Evans
Blue counter-stain. |

www.cdc.gov

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

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Table 5.1: Characteristics of the D FastPoint L-DFA Kit are compared to those of the following
	Diagnostic Hybrids (DHI) predicate devices
D3 FastPoint L-DFA	D3 Ultra Kit	D3 Duet RSV Kit	D' MPV Kit
Characteristics	Kit Subject Device	510(k) #K061101	510(k) # K081928	510(k) # K090073
	cytoplasmic and
punctate. Cells
appear round.
Parainfluenza 1, 2,
3: The fluorescence
is cytoplasmic. Cells
appear round.
Adenovirus: The
fluorescence is
cytoplasmic or
bright nuclear or
both. Cells appear
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.	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.	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)	l 3 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 (HINI),
WS (HIN1), PR
(H1N1), Wisconsin
(H3N2), WS
(HINI),
A/NWS/33 (HIN1)	10 influenza A
strains: Aichi
(H3N2), Mal
(H1N1), Hong Kong
(H3N2), Denver
(H1N1), Port
Chalmers (H3N2),
Victoria (H3N2),
New Jersey (H1N1),
WS (HINI),
PR,(HINI),
A/NWS/33 (H1N1)	10 influenza A
strains: Aichi
(H3N2), Mal
(H1N1), Hong Kong
(H3N2), Denver
(H1N1), Port
Chalmers (H3N2),
Victoria (H3N2),
New Jersey (HINI),
WS (HINI), PR
(HIN1), A/NWS/33
(HINI)	No reaction was
seen to any of the
tested influenza A
viruses with the
Metapneumovirus
DFA Reagent
Analytical specificity (for
Influenza B virus strains;
MAbs are reactive with all
listed strains)	7 influenza B
strains: Hong Kong,
Maryland, Mass,	7 influenza B
strains: Hong Kong,
Maryland, Mass,	7 influenza B
strains: Hong Kong,
Maryland, Mass,	No reaction was
seen to any of the
tested influenza B
Table 5.1: Characteristics of the D³ FastPoint L-DFA Kit are compared to those of the following .
Diagnostic Hybrids (DHI) predicate devices
Characteristics	D³ FastPoint L-DFA
Kit Subject Device	D³ Ultra Kit
510(k) #K061101	D³ Duet RSV Kit
510(k) # K081928	D³ MPV Kit
510(k) # K090073
	GL, Taiwan,
B/Lee/40, Russia	GL, Taiwan,
B/Lee/40, Russia	GL, Taiwan,
B/Lee/40, Russia	viruses with the
Metapneumovirus
DFA Reagent
Analytical
specificity
(cross-reactivity
studies; various
strains of
microorganisms
and cell lines)	Viruses	22	31	32	59
	Bacteria	22	18	25	25
	Chlamydia
spp.	1	1	3	3
	Yeast	1	0	1	1
	Protozoan	01	0	1	1
	Cell lines	N/A	17	17	16

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Although the D2 FastPoint L-DFA Influenza A/Influenza B Reagent has been shown to detect the 2009 HIN1 virus in two culture isolates, the performance characteristics of this device with clinical specimens that are positive for the 2009 HINI influenza virus have not been established. The D2 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 3 panels of proficiency-level antigen control slides. Each of the 3 reproducibility panels consisted 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.
- Mid level non-infected (negative) cells. e.

The RSV/hMPV panel consisted of the following:

a. Low level RSV (Washington strain) infected cells.
b. Low level hMPV (A1 subtype) infected cells.
c. Low level RSV (Washington strain) infected cells mixed with mid level hMPV (A1 subtype) infected cells.
d. Low level hMPV (A1 subtype) infected cells mixed with mid level RSV (Washington strain) infected cells.
Mid level non-infected (negative) cells. e.

10

The HPIV/Adenovirus panel consisted of the following:

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

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 x108 to 3.5 x108 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-yellow fluorescence.
c. Presence or absence of apple-green fluorescence.
d. Percent of cells exhibiting apple-green fluorescence.

For the D3 FastPoint L-DFAInfluenza A/Influenza B 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 FITC-labeled 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 D3 FastPoint L-DFAa A/Influenza B Reagent was 99.3% (278/280):

Table 5.2: Reproducibility Study Results using the D³ FastPoint L-DFA Influenza A/Influenza B Reagent
Panel
Member	Negative	Flu A
Low
Level	Flu B
Low
Level	Mixed Infection
Flu A Mid
Level	Mixed Infection
Flu B Low
Level	Mixed Infection
Flu A Low
Level	Mixed Infection
Flu B Mid
Level	Total %
Agreement
Concentration	No
infected
cells	4 to 10%
infected
cells	4 to 10%
infected
cells	20 to 30%
infected
cells	4 to 10%
infected
cells	4 to 10%
infected
cells	20 to 30%
infected
cells
Site 1
Agreement
with
Expected
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%)

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| Site
2 | Agreement
with
Expected
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%) |
|-----------|-----------------------------------------------|-----------------|-----------------|-----------------|-----------------|-----------------|-----------------|-----------------|--------------------|
| Site
3 | Agreement
with
Expected
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%) |
| Site
4 | Agreement
with
Expected
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
with Expected
result | 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% |

For the D3 FastPoint L-DFA RSV/hMPV Reagent, the combined data from the four Study Sites demonstrated reproducible detection of RSV by the R-PE labeled MAbs and reproducible detection of hMPV by the FITC-labeled MAbs. The presence of RSV infected cells was reported in 100% (120/120) of the wells in which the infected cells were expected. The presence of hMPV 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 RSV/hMPV Reagent was 100% (280/280):

		Table 5.3: Reproducibility Study Results using the D3 FastPoint L-DFA RSV/hMPV Reagent
	Panel
Member		RSV
Low
Level	hMPV
Low
Level	Mixed Infection		Mixed Infection
		Negative		Concentration	No
infected
cells	4 to 10%
infected
cells	4 to 10%
infected
cells	20 to 30%
infected
cells	4 to 10%
infected
cells
Site
1	Agreement
with
Expected
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%)
Site
2	Agreement
with
Expected
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%)
Site
3	Agreement
with
Expected
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%)

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D' FastPoint L-DFA Respiratory Virus Identification Kit 8/24/2009 Section 05, Page 13 of 23

| Site | Agreement
with
Expected
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%) |
|------|-----------------------------------------------|-----------------|-----------------|-----------------|-----------------|-----------------|-----------------|-----------------|-------------------|
| 4 | Total
Agreement
with Expected
result | 40/40
(100%) | 40/40
(100%) | 40/40
(100%) | 40/40
(100%) | 40/40
(100%) | 40/40
(100%) | 40/40
(100%) | 280/280
(100%) |
| | 95% CI | 91.2 -
100% | 91.2 -
100% | 91.2 -
100% | 91.2 -
100% | 91.2 -
100% | 91.2 -
100% | 91.2 -
100% | 98.7 -
100% |

For the D3 FastPoint L-DFA HPIV/Adenovirus Reagent, the combined data from the four Study Sites demonstrated reproducible detection of 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 HPIV/Adenovirus was 100% (280/280):

Table 5.4: Reproducibility Study Results using the D³ FastPoint L-DFA HPIV/Adenovirus Reagent
	Panel
Member	Negative	HPIV-1
Low Level	Adenovirus
Low Level	Mixed Infection		Mixed Infection	Total %
Agreement
	Concentration	No
infected
cells	4 to 10%
infected
cells	4 to 10%
infected
cells	HPIV-1
Mid Level	Adenovirus
Low Level	HPIV-1
Low Level	Adenovirus
Mid Level
Site
1	Agreement
with
Expected
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%)
Site
2	Agreement
with
Expected
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%)
Site
3	Agreement
with
Expected
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%)
Site
4	Agreement
with
Expected
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
with Expected
result	40/40
(100%)	40/40
(100%)	40/40
(100%)	40/40
(100%)	40/40
(100%)	40/40
(100%)	40/40
(100%)	280/280
(100%)

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	Table 5.4:	Reproducibility Study Results using the D³ FastPoint L-DFA HPIV/Adenovirus Reagent
95% CI	91.2 – 100%	91.2 – 100%	91.2 – 100%	91.2 – 100%	91.2 – 100%	91.2 – 100%	98.7 – 100%

Limit of Detection

Analytical Limit of Detections (LoDs) of the D3 FastPoint L-DFA Reagents was addressed using dilution series of infected model cells. Model cells for 8 characterized respiratory virus isolates; influenza A virus (ATCC Victoria strain), influenza B virus (ATCC Taiwan strain), respiratory syncytial virus (ATCC Washington strain), adenovirus (ATCC type 1), human metapneumovirus subtype A1 (clinical strain), parainfluenza virus types 1, 2, and 3 (ATCC strains C-35, Greer, and C243 respectively) 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 25uL 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 snot 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 5.5 below:

| Table 5.5:

Kit	Limit of Detections of the D3 FastPoint L-DFA Respiratory Virus Identification
Virus Strain	Infected cells/mL	Number of replicates with positive cells	LOD determination
	500	10/10
	100	10/10
	50	10/10
	25	5/10
Flu A
(ATCC Victoria strain)	12.5	3/10	50 infected cells/mL
	6	2/10
	3	0/10
	1.5	2/10
	0.8	0/10
	0.4	0/10
	2000	10/10
	400	10/10
	200	10/10
	100	10/10
Flu B
(ATCC Taiwan strain)	50	10/10	50 infected cells/mL
	25	7/10
	12.5	4/10
	6	2/10
	3	0/10
	1.5	0/10
Table 5.5:
Kit		Limit of Detections of the D³ FastPoint L-DFA Respiratory Virus Identification
Virus Strain	Infected cells/mL	Number of replicates with
positive cells	LOD determination
	1000	10/10
	200	10/10
	100	10/10
	50	7/10
RSV
(ATCC Washington
strain)	25	7/10	100 infected cells/mL
	12.5	6/10
	6	1/10
	3	0/10
	1.5	0/10
	0.8	0/10
	2000	10/10
	400	10/10
	200	10/10
	100	10/10
hMPV A1
(Clinical strain)	50	6/10	100 infected cells/mL
	25	2/10
	12.5	0/10
	6	0/10
	3	0/10
	1.5	0/10
	1000	10/10
	200	10/10
	100	9/10
	50	5/10
Adenovirus
(ATCC type 1)	25	1/10	100 infected cells/mL
	12.5	0/10
	6	0/10
	3	0/10
	1.5	0/10
	0.8	0/10
	500	10/10
	100	10/10
	50	6/10
	25	2/10
HPIV-1
(ATCC strain C-35)	12.5	1/10	100 infected cells/mL
	6	0/10
	3	0/10
	1.5	0/10
	0.8	0/10
	0.4	0/10
	500	10/10
	100	10/10
	50	10/10
	25	9/10
	HPIV-2
(ATCC strain Greer)	12.5	6/10
(ATCC strain Greer)	6	5/10
	3	3/10
	1.5	1/10
	0.8	0/10
	0.4	0/10
HPIV-3
(ATCC strain C243)	1000	10/10	50 infected cells/mL
	200	10/10
	100	10/10
	50	9/10
Table 5.5: Limit of Detections of the D3 FastPoint L-DFA Respiratory Virus Identification
Kit
Virus Strain	Infected cells/mL	Number of replicates with
positive cells	LOD determination
	25	6/10
	12.5	2/10
	6	0/10
	3	0/10
	1.5	0/10
	0.8	0/10

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Analytical reactivity (inclusivity)

Analytical reactivity (inclusivity) of the D3FastPoint L-DFA Influenza A/Influenza B 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 D3 FastPoint L-DFA Influenza A/Influenza B Reagent.

| Table 5.6: Analytical Reactivity (inclusivity) of the D³ FastPoint L-DFA Influenza A/Influenza B

Reagent on various influenza A virus and influenza B virus strains
Influenza Strains	Infected Cell Concentration
(as multiples of the respective
established LoD concentration	D³ FastPoint L-DFA Influenza A/
Influenza B 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

Sec05_FastPoint_L-DFA_Final.doc

16

Diagnostic Hybrids, Inc.

D3 FastPoint L-DFA Respiratory Virus Identification Kit 8/24/2009 Section 05, Page 17 of 23

| Table 5.6: Analytical Reactivity (inclusivity) of the D3, FastPoint L-DFA Influenza A/Influenza B

Reagent on various influenza A virus and influenza B virus strains
Influenza Strains	Infected Cell Concentration
(as multiples of the respective
established LoD concentration	D3 FastPoint L-DFA Influenza A/
Influenza B Reagent Results
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 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 reactivity (inclusivity) of the D3 FastPoint L-DFA RSV/hMPV DFA Reagent was evaluated using 3 RSV virus and 4 hMPV 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 D FastPoint L-DFA RSV/hMPV Reagent.

| Table 5.7: Analytical Reactivity (inclusivity) of the D3 FastPoint L-DFA RSV/hMPV DFA

Reagent on various RSV virus and hMPV virus strains
RSV and hMPV Strains	Infected Cell Concentration (as
multiples of the respective
established LoD concentration	D3 FastPoint L-DFA RSV/hMPV Reagent
Results
RSV 9320	10x LoD	22 Golden-yellow fluorescent cells
RSV Washington	10x LoD	22 Golden-yellow fluorescent cells
RSV Long	10x LoD	32 Golden-yellow fluorescent cells
hMPV A1	10x LoD	25 Apple-green fluorescent cells
hMPV A2	10x LoD	25 Apple-green fluorescent cells
hMPV B1	10x LoD	25 Apple-green fluorescent cells
hMPV B2	10x LoD	37 Apple-green fluorescent cells

Analytical reactivity (inclusivity) of the D3 FastPoint L-DFA HPIV/Adenovirus DFA Reagent was evaluated using 3 HPIV virus and 10 Adenovirus 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 D FastPoint L-DFA HPIV/Adenovirus Reagent.

17

| Table 5.8: | Analytical Reactivity (inclusivity) of the D³ FastPoint L-DFA HPIV/Adenovirus
Reagent on various HPIV virus and adenovirus strains | |
|--------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------|
| Parainfluenza and Adenovirus Strains | Infected Cell Concentration
(as multiples of the respective
established LoD
concentration | D³ FastPoint L-DFA HPIV/Adenovirus
Reagent Results |
| Parainfluenza 1 C-35 | 10x LoD | 9 Golden-yellow fluorescent cells |
| Parainfluenza 2 Greer | 10x LoD | 11 Golden-yellow fluorescent cells |
| Parainfluenza 3 C-243 | 10x LoD | 22 Golden-yellow fluorescent cells |
| Adenovirus 1 VR-1 | 10x LoD | 26 Apple-green fluorescent cells |
| Adenovirus 3 VR-3 | 10x LoD | 17 Apple-green fluorescent cells |
| Adenovirus 5 VR-5 | 10x LoD | 15 Apple-green fluorescent cells |
| Adenovirus 6 VR-6 | 10x LoD | 22 Apple-green fluorescent cells |
| Adenovirus 7 VR-7 | 10x LoD | 16 Apple-green fluorescent cells |
| Adenovirus 8 VR-1366 | 10x LoD | 29 Apple-green fluorescent cells |
| Adenovirus 10 VR-1087 | 10x LoD | 34 Apple-green fluorescent cells |
| Adenovirus VR-14 | 10x LoD | 37 Apple-green fluorescent cells |
| Adenovirus Dewitt ATCC Strain | 10x LoD | 15 Apple-green fluorescent cells |
| Adenovirus 31 VR-1109 | 10x LoD | 42 Apple-green fluorescent cells |

Clinical Performance:

Performance of the D3 FastPoint L-DFA Respiratory Virus Identification 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 L-DFA Respiratory Virus Identification Kit was assessed and compared to a predetermined algorithm that used composite comparator methods. The composite comparator methods for influenza A virus, influenza B virus, respiratory syncytial virus, 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). For human metapneumovirus the composite comparator methods consisted of DSFA with an FDA cleared device, and confirmation of all negative specimens (as determined by the comparator

18

DSFA test) using a validated3 hMPV real-time RT-PCR followed by bidirectional sequencing analysis comparator assay. The hMPV real-time RT-PCR comparator assay targets the hMPV Nucleocapsid gene. "True" positive was defined as any sample that either tested positive by the comparator DSFA test or viral culture, or had bi-directional sequencing data meeting pre-defined quality acceptance criteria that matched hMPV sequences deposited in the National Center for Biotechnology Information (NCBI) GenBank database (www.ncbi.nlm.nih.gov), with acceptable E-values4 "True" negative was defined as any sample that tested negative by both the comparator DSFA test and either viral culture or the hMPV real-time RT-PCR comparator assay.

Prevalence of the respiratory viruses within this population as determined by the D3 FastPoint L-DFA Respiratory Virus Identification Kit direct specimen testing is noted in Table 5.9 below:

Table 5.9: Respiratory Virus Prevalence
Age	Total
Specimens
Evaluated	Flu A

positive

(prevalence) | Flu B

positive

(prevalence) | RSV

positive

(prevalence) | hMPV

positive

(prevalence) | Adenovirus

positive

(prevalence) | HPIV

positive

(prevalence) |
| 0 - 1
month | 55 | 0 | 0 | 15 (27.3%) | 2 (3.6%) | 0 | 1 (1.8%) |
| > 1
month to
2 years | 577 | 27 (4.7%) | 20 (3.5%) | 154 (26.7%) | 41 (7.1%) | 11 (1.9%) | 29 (5.0%) |
| > 2
years to
12 years | 391 | 43 (11.0%) | 104 (26.6%) | 25 (6.4%) | 17 (4.3%) | 1 (0.3%) | 6 (1.5%) |
| > 12
years to
21 years | 173 | 19 (11.0%) | 41 (23.7%) | 4 (2.3%) | 3 (1.7%) | 0 | 2 (1.2%) |
| 22 years
to 30
years | 57 | 3 (5.3%) | 14 (24.6%) | 0 | 1 (1.8%) | 0 | 1 (1.8%) |
| 31 years
to 40
years | 71 | 9 (12.7%) | 9 (12.7%) | 1 (1.4%) | 3 (4.2%) | 0 | 0 |
| 41 years
to 50
years | 52 | 5 (9.6%) | 5 (9.6%) | 0 | 1 (1.9%) | 0 | 0 |

Analytical validation of the real-time hMPV RT-PCR followed by bi-directional sequencing analysis comparator assay included analytical sensitivity and reactivity study, analytical specificity study, and extraction efficiency study. The analytical sensitivity (limit of detection or LoD) of the real-time hMPV RT-PCR followed by bi-directional sequencing analysis comparator assay was determined using quantified (TCID30mL) stocks of the 4 hMPV (subtypes A1, A2, B1 and B2) strains diluted in hMPV negative nasopharyngeal clinical matrix, and ranged from 10 - 50 TCIDs/mL.

4 The E-values generated from the clinical trials range from a low of 5e-78 to a high of 1e-20. The E-Value from NCBI BLAST Alignment indicates the statistical significance of a given pair-wise alignment and reflects the size of the database and the scoring system used. The lower the E-Value, the more significant the hit. A sequence alignment that has an E-Value of 1e-3 means that this similarity has a 1 in 1000 chance of occurring by chance alone.

(http://www.ncbi.nlm.nih.gov/books/bv.fcgi?rid=handbook.section.614).

Sec05 FastPoint L-DFA Final.doc

19

	Table 5.9:		Respiratory Virus Prevalence
Age	Total
Specimens
Evaluated	Flu A	Flu B	RSV	hMPV	Adenovirus	HPIV
		# positive
(prevalence)	# positive
(prevalence)	# positive
(prevalence)	# positive
(prevalence)	# positive
(prevalence)	# positive
(prevalence)
51 years
to 60
years	46	3 (6.5%)	3 (6.5%)	1 (2.2%)	3 (6.5%)	0	0
61 years
to 70
years	33	2 (6.1%)	2 (6.1%)	1 (3.0%)	1 (3.0%)	0	0
71 years
to 80
years	16	2 (12.5%)	1 (6.3%)	1 (6.3%)	4 (25.0%)	0	0
81 years
and
above	7	0	0	1 (14.3%)	0	0	1 (14.3%)
Age Not
Reported	41	2 (4.9%)	14 (34.1%)	0	1 (2.4%)	1 (2.4%)	0
Total	1519	115 (7.6%)	213 (14.0%)	203 (13.4%)	77 (5.1%)	13 (0.9%)	40 (2.6%)

There were seven (7) co-infections detected: 2 - respiratory syncytial virus + metapneumovirus, 2- adenovirus + respiratory syncytial virus, 2- influenza A + metapneumovirus, 1-respiratory syncytial virus + adenovirus and 1-respiratory syncytial virus + parainfluenza virus

Tables 5.10 through 5.15 below show the study results of the NP wash/aspirate specimen type (Sites 1, 2, and 3 combined):

Table 5-10: Flu A
Fresh nasal/nasopharyngeal wash/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 5.1: Flu B
Fresh nasal/nasopharyngeal
Of limited use, difficult to interpret, and low sensitivity

wash/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%

Sec05_FastPoint_L-DFA_Final.doc

20

D³ FastPoint L-DFA Respiratory Virus Identification Kit

Section 05, Page 21 of 23

| Fresh nasal/nasopharyngeal
wash/aspirate | Comparator DSFA (negatives followed by culture with
DFA) | | |
|---------------------------------------------|-------------------------------------------------------------|----------|---------------|
| DHI DSFA | Positive | Negative | Total |
| Positive | 204 | 1 | 205 |
| Negative | 3 | 462 | 465 |
| Total | 207 | 463 | 670 |
| | | | 95% CI |
| Sensitivity | 204/207 | 98.6% | 95.8-99.7% |
| Specificity | 462/463 | 99.8% | 98.8-100% |

| ST . BRPT. 122
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|----------------------------------------------------------------------------------------------------------|--------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| 1000 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 100 - 1
STATE CHARE START CO | | |

| Fresh nasal/nasopharyngeal
wash/aspirate | Comparator DSFA (negatives followed by culture with
DFA) | | |
|---------------------------------------------|-------------------------------------------------------------|----------|---------------|
| DHI DSFA | Positive | Negative | Total |
| Positive | 12 | 0 | 12 |
| Negative | 1 | 619 | 620 |
| Total | 13 | 619 | 632 |
| | | | 95% CI |
| Sensitivity | 12/13 | 92.3% | 64.0-99.8% |
| Specificity | 619/619 | 100.0% | 99.4-100% |

Table 5.14: HPIV
Fresh nasal/nasopharyngeal
wash/aspirate	Comparator DSFA (negatives followed by culture with
DFA)
DHI DSFA	Positive	Negative	Total
Positive	23	4	27
Negative	2	599	601
Total	25	603	628
			95% CI
Sensitivity	23/25	92.0%	74.0-99.0%
Specificity	599/603	99.3%	98.3-99.8%

Table 5.15: Table 5.15: Table 5.15: hMPV Sale Sale Sale ﺍﻟﻤﺴﺘﻘﻠﺔ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘ ﺍﻟﻤﻮﺍﻗﻊ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘﻮﻯ ﺍﻟﻤﺴﺘ

| Fresh nasal/nasopharyngeal
wash/aspirate | Comparator DSFA (negatives confirmed by a validated
hMPV real-time RT-PCR followed by bi-directional
sequencing analysis comparator assay) | | |
|---------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------|----------|------------|
| DHI DSFA | Positive | Negative | Total |
| Positive | 55 | 0 | 55 |
| Negative | 25 | 614 | 639 |
| Total | 80 | 614 | 694 |
| Sensitivity | 55/80 | 68.8% | 57.4-78.7% |
| Specificity | 614/614 | 100.0% | 99.4-100% |

21

Tables 5.16 through 5.21 below show the study results of the NP swab specimen type (Sites 3 and 4 combined):

Table 5.16: Flu A
Fresh nasal/nasopharyngeal swab	Comparator DSFA (negatives followed by culture with
DFA)
DHI DSFA	Positive	Negative	Total
Positive	57	1	58
Negative	8	623	631
Total	65	624	689
			95% CI
Sensitivity	57/65	87.7%	77.2-94.5%
Specificity	623/624	99.8%	99.1-100%

Table 5.17: Flu B.
Fresh nasal/nasopharyngeal swab	Comparator DSFA (negatives followed by culture with DFA)
DHI DSFA	Positive	Negative	Total
Positive	203	1	204
Negative	28	478	506
Total	231	479	710
			95% CI
Sensitivity	203/231	87.9%	83.7-92.1%
Specificity	478/479	99.8%	98.8-100%

Table 5.18: RSV
Fresh nasal/nasopharyngeal swab	Comparator DSFA (negatives followed by culture with DFA)
DHI DSFA	Positive	Negative	Total
Positive	39	0	39
Negative	1	646	647
Total	40	646	686
			95% CI
Sensitivity	39/40	97.5%	86.8-99.9%
Specificity	646/646	100.0%	99.4-100%

	Table 5.19:	Adenovirus
Fresh nasal/nasopharyngeal swab	Comparator DSFA (negatives followed by culture with
DFA)
DHI DSFA	Positive	Negative	Total
Positive	1	0	1
Negative	0	679	679
Total	1	679	680

Sec05_FastPoint_L-DFA_Final.doc

.

22

8/24/2009 Section 05, Page 23 of 23

			95% CI
Sensitivity	1/1	100.0%	NA
Specificity	679/679	100.0%	99.5-100%

Note: The sensitivity performance of the D FastPoint L-DFA Respiratory Virus ID 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.

	Table 5.20:	HPIV
Fresh nasal/nasopharyngeal swab	Comparator DSFA (negatives followed by culture with
DFA)
DHI DSFA	Positive	Negative	Total
Positive	13	0	13
Negative	1	667	668
Total	14	667	681
			95% CI
Sensitivity	13/14	92.9%	66.1-99.8%
Specificity	667/667	100.0%	99.4-100%

		Table 5.21: hMPV
Fresh nasal/nasopharyngeal swab	Comparator DSFA (negatives confirmed by a validated
hMPV real-time RT-PCR followed by bi-directional
sequencing analysis comparator assay)
DHI DSFA	Positive	Negative	Total
Positive	24	0	24
Negative	20	631	651
Total	44	631	675
			95% CI
Sensitivity	24/44	54.5%	38.8-69.9%
Specificity	631/631	100.0%	99.4-100%

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

23

Image /page/23/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES. USA" around the perimeter. Inside the circle is a stylized eagle-like symbol with three curved lines representing its wings or feathers.

DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

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

Mr. Ronald Lollar

SEP 112009

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

Re: K091171

Trade/Device Name: D3 FastPoint L-DFA Respiratory Virus Identification Kit Regulation Number: 21 CFR 866.3980

Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: Class II

Product Code: OMG, LKT, GNX, GOS, GNY

Dated: September 2, 2009

Received: September 8, 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

24

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.

Ube Scrf. fir

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

25

Indications for Use

510(k) Number (if known): K091171

Device Name: D FastPoint L-DFA Respiratory Virus Identification Kit

Indications For Use:

The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA Respiratory Virus Identification Kit is intended for the qualitative identification of influenza A virus, influenza B virus, respiratory syncytial virus, human metapneumovirus, 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 influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus or parainfluenza viruses after examination of the direct specimen result be confirmed by cell culture. Specimens found to be negative for human metapneumovirus after examination of the direct specimen results should be confirmed by an FDA cleared human metapneumovirus molecular assay. 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 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.2

Prescription Use X (Part 21 CFR 801 Subpart D) AND/OR

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

ﺎ ﺍﻟﻤﺮﺍﺟﻊ

Division Sign-Off

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

O(K) ki 1171

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

www.cdc.gov

26

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