Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description and performance studies focus on traditional immunofluorescence techniques and manual microscopic examination, with no mention of automated image analysis, AI, or ML.

Therapeutic

No.
This device is a diagnostic tool intended for the qualitative identification of specific viruses, not for providing therapy or treatment.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the device "is intended for the qualitative identification of respiratory syncytial virus and human metapneumovirus..." and also notes that "Negative results do not preclude respiratory syncytial virus and human metapneumovirus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions," indicating its role in diagnosis.

SaMD (Software as a Medical Device)

No

The device is a diagnostic kit that includes reagents (monoclonal antibodies) and requires the use of a fluorescence microscope for examination, indicating it is a hardware-based medical device, not software-only.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the device is "intended for the qualitative identification of respiratory syncytial virus and human metapneumovirus 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)." This clearly describes a test performed on specimens taken from the human body to provide information for diagnosis.
Device Description: The description details a process of using reagents (monoclonal antibodies) to stain cells from patient specimens and then examining these stained cells using a fluorescence microscope to identify the presence of specific viral antigens. This is a typical in vitro diagnostic procedure.
Specimen Type: The device uses "nasal and nasopharyngeal swabs and aspirates/washes specimens," which are specimens taken from the human body.
Purpose: The purpose is the "qualitative identification" of specific viruses, which is a diagnostic activity.

All of these points align with the definition of an In Vitro Diagnostic device, which is a medical device intended for use 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 RSV/MPV Identification Kit is intended for the qualitative identification of respiratory syncytial virus and human metapneumovirus 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 respiratory syncytial virus 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 syncytial virus and human metapneumovirus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.

Product codes

OMG, LKT

Device Description

The D3 FastPoint L-DFA RSV/MPV Identification Kit uses a blend (called a "L-DFA Reagent'") of viral antigen-specific murine monoclonal antibodies that are directly labeled with either R-phycoerythin (PE) (respiratory syncytial virus) or fluorescein isothiocyanate (FITC) (human metapneumovirus) for the rapid identification of respiratory syncytial virus and human metapneumovirus in nasal and nasopharyngeal swabs and aspirates from patients with signs and symptoms of respiratory infection.

Kit Components:

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 human metapneumovirus 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 RSV/MPV 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 respiratory syncytial virus or human metapneumovirus. The negative wells contain non-infected cells. Each slide is intended to be stained only one time.
D3 FastPoint L-DFA Specimen Slides and Coverslips, 50-slides with coverslips. Fifty pack of 3-well specimen slides.

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

Description of the test set: Clinical Performance studies used 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.
Sample size: 1519 total specimens evaluated across all ages. Specific specimen types: 670 fresh nasal/nasopharyngeal wash/aspirate specimens, 687 fresh nasal/nasopharyngeal swab specimens.
Data source: 4 geographically diverse U.S. clinical laboratories during the 2009 respiratory virus seasons (January 2009 - March 2009).
Annotation protocol: 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. The D3 FastPoint RSV/MPV Kit was assessed and compared to a predetermined algorithm that used composite comparator methods.
For respiratory syncytial virus: 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: DSFA with an FDA-cleared device, and confirmation of all negative specimens (as determined by the comparator DSFA test) using a validated hMPV real-time RT-PCR followed by bi-directional sequencing analysis comparator assay.
"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-values. "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.

Summary of Performance Studies

Study type: Analytical Performance studies (Precision/Reproducibility, Limit of Detection, Analytical reactivity) and Clinical Performance studies.
Sample size:
Precision/Reproducibility: 5 randomized panel members tested daily in two separate runs for 5 days by four different laboratories (40 total runs). Each sample contains 2.5 x 10^5 to 3.5 x 10^5 total cells.
Limit of Detection: Dilution series of infected model cells, 10 replicates for each dilution level.
Analytical Reactivity: Low concentration infected cell suspensions (approximately 4% cells infected, 25 to 50 infected cells) for 3 RSV virus and 4 hMPV virus strains.
Clinical Performance: 1519 total specimens evaluated. Specific analysis on 670 fresh nasal/nasopharyngeal wash/aspirate specimens and 687 fresh nasal/nasopharyngeal swab specimens.

AUC: Not Found
MRMC: Not Found
Standalone performance: Not Found

Key results:
Precision/Reproducibility: 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 L-DFA Reagent was 100% (280/280).
Limit of Detection: For RSV, LoD was 100 infected cells/mL. For hMPV A1, LoD was 100 infected cells/mL.
Analytical Reactivity: All tested RSV and hMPV strains showed positive fluorescent cells when stained with the L-DFA Reagent at 10x LoD.
Clinical Performance (RSV - NP wash/aspirate): Sensitivity 98.6% (204/207), Specificity 99.8% (462/463).
Clinical Performance (hMPV - NP wash/aspirate): Sensitivity 68.8% (55/80), Specificity 100.0% (614/614).
Clinical Performance (RSV - NP swab): Sensitivity 97.5% (39/40), Specificity 100.0% (647/647).
Clinical Performance (hMPV - NP swab): Sensitivity 54.5% (24/44), Specificity 100.0% (632/632).
Overall, devices detect RSV and hMPV antigens in a similar manner.

Key Metrics

For NP wash/aspirate specimens:
RSV:
Sensitivity: 98.6% (95% CI: 95.8-99.7%)
Specificity: 99.8% (95% CI: 98.8-100%)
hMPV:
Sensitivity: 68.8% (95% CI: 57.4-78.7%)
Specificity: 100.0% (95% CI: 99.4-100%)

For NP swab specimens:
RSV:
Sensitivity: 97.5% (95% CI: 86.8-99.9%)
Specificity: 100.0% (95% CI: 99.4-100%)
hMPV:
Sensitivity: 54.5% (95% CI: 38.8-69.9%)
Specificity: 100.0% (95% CI: 99.4-100%)

Predicate Device(s)

K061101, K081928, 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

1093233

Diagnostic Hybrids, Inc.

D3 FastPoint L-DFA RSV/MPV Identification Kit

10/06/2009 Page 1 of 14

Section 05, 510(k) Summary

Applicant:

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

Contact Information:

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

Date of preparation of 510(k) summary:

October 5. 2009

Device Name:

Trade name - D FastPoint L-DFA RSV/MPV Identification Kit Common name - RSV/MPV DFA assay Classification name - Respiratory viral panel multiplex nucleic acid assay Product Code - OMG, LKT Regulation - 21 CFR 866.3980 Regulatory Class - Class II 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

DEC - 4 2009

1

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

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.

D' Duet DFA RSV/Respiratory Virus Screening Kit (K081928)

Intended Use: 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

2

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 BSL3+ facility is available to receive and culture specimens.

D3 DFA Metapneumovirus Identification Kit (K090073)

Intended Use: The Diagnostic Hybrids, Inc. device, D3 DFA Metapneumovirus Identification Kit (D3 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 FDA-cleared hMPV molecular assay.

Device Description:

The D3 FastPoint L-DFA RSV/MPV Identification Kit uses a blend (called a "L-DFA Reagent'") of viral antigen-specific murine monoclonal antibodies that are directly labeled with either R-phycoerythin (PE) (respiratory syncytial virus) or fluorescein isothiocyanate (FITC) (human metapneumovirus) for the rapid identification of respiratory syncytial virus and human metapneumovirus in nasal and nasopharyngeal swabs and aspirates from patients with signs and symptoms of respiratory infection.

Kit Components:

D3 FastPoint L-DFA RSV/MPV Reagent, 4.0-mL. One dropper bottle 1. containing a mixture of PE-labeled murine monoclonal antibodies directed against respiratory syncytial virus antigens and FITC-labeled murine monoclonal antibodies directed against human metapneumovirus antigens. The buffered, stabilized, aqueous solution contains Evans Blue and propidium iodide as counter-stains and 0.1% sodium azide as preservative.
40X PBS Concentrate, 25-mL. One bottle of 40X PBS concentrate containing 2. 4% sodium azide (0.1% sodium azide after dilution to 1X using de-mineralized water).

3

Re-suspension Buffer, 6.0-mL. One bottle of a buffered glycerol solution and 3. 0.1% sodium azide.
D3 FastPoint L-DFA RSV/MPV Antigen Control Slides, 5-slides. Five 4. individually packaged control slides containing 2 wells with cell culture-derived positive and negative control cells. Each positive well contains cells infected with either respiratory syncytial virus or human metapneumovirus. The negative wells contain non-infected cells. Each slide is intended to be stained only one time.
D3 FastPoint L-DFA Specimen Slides and Coverslips, 50-slides with 5. coverslips. Fifty pack of 3-well specimen slides.

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

Intended Use:

The Diagnostic Hybrids, Inc. device, D3 FastPoint L-DFA RSV/MPV Identification Kit is intended for the qualitative identification of respiratory syncytial virus and human metapneumovirus 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 respiratory syncytial virus 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 syncytial virus and human metapneumovirus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions. ·

4

·

| 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
RSV/MPV
Identification 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
RSV/MPV
Identification Kit is
intended for the
qualitative
identification of
respiratory syncytial
virus and human
metapneumovirus 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 respiratory
syncytial virus 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	The Diagnostic
Hybrids, Inc. D3
UltraTM DFA (direct
fluorescent
antibody)
Respiratory Virus
Screening & ID Kit
is intended for the
qualitative detection
and identification of
the influenza A,
influenza B,
respiratory syncytial
virus (RSV),
adenovirus,
parainfluenza 1,
parainfluenza 2 and
parainfluenza 3
virus in respiratory
specimens, by either
direct detection or
cell culture method,
by
immunofluorescence
using 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.	The Diagnostic
Hybrids, Inc. device,
D3 Duet DFA
RSV/Respiratory
Virus Screening Kit,
is intended for the
qualitative detection
and identification of
respiratory syncytial
virus, while
screening for
influenza A virus,
influenza B virus,
adenovirus, and
parainfluenza virus
types 1, 2 and 3 viral
antigens, in nasal
and nasopharyngeal
swabs and aspirates
or in cell culture.
The assay detects
viral antigens by
immunofluorescence
using monoclonal
antibodies (MAbs),
from patients with
signs and symptoms
of respiratory
infection.
It is recommended
that specimens
found to be negative
after examination of
the direct specimen
result be confirmed
by cell culture.
Negative results do
not preclude
influenza virus .
infection and should
not be used as the
sole basis for
diagnosis, treatment
or other
management	The Diagnostic
Hybrids, Inc. device,
D3 DFA
Metapneumovirus
Identification 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
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
RSV/MPV
Identification Kit
(Subject Device)	D3 Ultra Kit
510(k) #K061101	D3 Duet RSV Kit
510(k) # K081928	D3 MPV Kit
510(k) # K090073
	not preclude
respiratory virus
infection and should
not be used as the
sole basis for
diagnosis, treatment
or other
management
decisions.		decisions.	results be confirmed
by an FDA-cleared
hMPV molecular
assay.
Target Viruses	respiratory syncytial
virus,
metapneumovirus	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 RSV/MPV
Reagent contain 5
MAbs to respiratory
syncytial virus (2)
and
metapneumovirus
(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
MAbs to RSV. | Direct labeling
using fluorescein
isothiocyanate
(FITC) to label all
MAbs with
fluorescein. | Direct labeling
using R-
Phycoerythrin (R-
PE) to label the
MAbs to respiratory
syncytial virus.
using fluorescein
isothiocyanate | Direct labeling
using fluorescein
isothiocyanate
(FITC) to label all
MAbs with
fluorescein. | |
| | | | | | |
| 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
RSV/MPV
Identification Kit
(Subject Device) | D³ Ultra Kit
510(k) #K061101 | D³ Duet RSV Kit
510(k) #K081928 | D³ MPV Kit
510(k) # K090073 | |
| | (FITC) to label the
MAbs to
metapneumovirus. | | (FITC) to label all
other MAbs with
fluorescein. | | |
| R-Phycoerythrin-labeled
MAbs | respiratory syncytial
virus | None | respiratory syncytial
virus | None | |
| Fluorescein-labeled MAbs | metapneumovirus | influenza A virus,
influenza B virus,
respiratory syncytial
virus, adenovirus,
parainfluenzavirus
type 1,
parainfluenzavirus
type 2,
parainfluenzavirus
type 3 | influenza A virus,
influenza B virus,
adenovirus,
parainfluenzavirus
type 1, parainfluenza
virus type 2,
parainfluenzavirus
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 | Respiratory
Syncytial Virus:
The fluorescence is
cytoplasmic. Cells
appear round.
Metapneumovirus:
The fluorescence is
cytoplasmic and
punctate. 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. | Influenza A and B:
The fluorescence is
cytoplasmic, nuclear
or both.
Cytoplasmic
staining is often
punctate with large
inclusions while
nuclear staining is
uniformly bright.
Respiratory
Syncytial Virus:
The fluorescence is
cytoplasmic and
punctate with small
inclusions in the
syncytia.
Parainfluenza 1, 2,
3: The fluorescence
is cytoplasmic and
punctate with
irregular inclusions.
Types 2 and 3 cause
the formation of
syncytia.
| Influenza A and
B: The
fluorescence is
cytoplasmic,
nuclear or both.
Cytoplasmic
staining is often
punctate with large
inclusions while
nuclear staining is
uniformly bright.
Respiratory
Syncytial Virus:
The fluorescence
is cytoplasmic and
punctate with
small inclusions in
the syncytia.
Parainfluenza 1,
2, 3: The
fluorescence is
cytoplasmic and
punctate with
irregular
inclusions. Types
| 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. | |
| 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
RSV/MPV
Identification Kit
(Subject Device) | D³ Ultra Kit
510(k) #K061101 | D³ Duet RSV Kit
510(k) # K081928 | D³ MPV Kit
510(k) # K090073 | |
| | | fluorescence is
cytoplasmic and
punctate or bright
nuclear or both.
Negative: Cells
fluoresce red due to
the Evans Blue
counter-stain. | 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. | | |
| Device Reagents are not reactive with these numbers of microorganisms. | | | | | |
| Analytical
specificity
(cross-reactivity
studies; various
strains of
microorganisms
and cell lines) | Viruses | 59 | 31 | 32 | 59 |
| | Bacteria | 22 | 18 | 25 | 25 |
| | Chlamydia
spp. | 1 | 1 | 3 | 3 |
| | Yeast | 1 | 0 | 1 | 1 |
| | Protozoan | 0 | 0 | 1 | 1 |
| | Cell lines | N/A | 17 | 17 | 16 |

Technological Characteristics, Compared to Predicate Device:

5

.

6

7

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 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.
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 x106 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.
c. Presence or absence of apple-green fluorescence.
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 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 L-DFA Reagent was 100% (280/280):

TABLE 5.2: Reproducibilty Study Results using the L-DFA Reagent
Site	Panel
Member	Negative	RSV
Low
Level	hMPV
Low
Level	Mixed Infection		Mixed Infection
				RSV Mid
Level	hMPV
Low Level	RSV
Low
Level	hMPV
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	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%)
	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%

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 respiratory syncytial virus (ATCC Washington strain) and human metapneumovirus

9

10/06/2009

subtype A1 (clinical strain) were diluted with non-infected cells to produce a suspension equivalent to 1,000 infected cells per milliliter. This level theoretically yields approximately 25 infected cells per 25-uL of suspension. This suspension was then serially diluted to a theoretical level of less than 1 cell per milliliter. (NOTE: This level was the target to begin with a low positive level. Actual starting levels vary, however, and are within 1 dilution of the 25 infected cell target level). 25-uL aliquots from each dilution level were spotted onto 10 replicate microscope slides, and then stained according to the instructions for use described in this product insert. Each cell spot was examined at 200x magnification. Results were reported as numbers of positive replicates for each set of 10. Analytical detection limits for each of the 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
RSV
(ATCC Washington
strain)	1000	10/10	100 infected cells/mL
	200	10/10
	100	10/10
	50	7/10
	25	7/10
	12.5	6/10
	6	1/10
	3	0/10
	1.5	0/10
	0.8	0/10
hMPV A1
(Clinical strain)	2000	10/10	100 infected cells/mL
	400	10/10
	200	10/10
	100	10/10
	50	6/10
	25	2/10
	12.5	0/10
	6	0/10
	3	0/10
	1.5	0/10

Analytical reactivity (inclusivity):

Analytical reactivity (inclusivity) of the L-DFA Reagent was evaluated using 3-RSV virus and 4-hMPV virus strains. Low concentration infected cell suspensions (approximately 4% cells infected, 25 to 50 infected cells) were prepared for each viral strain. The suspensions were stained with the L-DFA Reagent.

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TABLE 5.4: Analytical Reactivity (inclusivity) of the L-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)	L-DFA 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

Clinical Performance:

Performance of the Dr FastPoint RSV/MPV 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 RSV/MPV Kit was assessed and compared to a predetermined algorithm that used composite comparator methods. The composite comparator methods for respiratory syncytial virus 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 DSFA test) using a validated ' hMPV real-time RT-PCR followed by bi-directional sequencing analysis comparator assay. The hMPV real-time RT-PCR comparator assay targets the hMPV Nucleocapsid gene. "True"

Sec05 FastPoint RSV MPV 09-10061 FDA.doc

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 (TCIDs /mL) stocks of the 4 hMPV (subtypes A1, A2, B1 and B2) strains diluted in hMPV negative nasopharyngeal clinical matrix, and ranged from 10 - 50 TCID30mL.

11

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-values.2 "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 RSV and hMPV within this population as determined by the D3 FastPoint RSV/MPV Kit direct specimen testing is noted in TABLE 5.5 below:

TABLE 5.5: RSV/hMPV Prevalence*
Age	Total
Specimens
Evaluated	RSV

positive

(prevalence) | hMPV

positive

(prevalence) |
| 0 - 1 month | 55 | 15 (27.3%) | 2 (3.6%) |
| > 1 month to 2 years | 577 | 154 (26.7%) | 41 (7.1%) |
| > 2 years to 12 years | 391 | 25 (6.4%) | 17 (4.3%) |
| > 12 years to 21 years | 173 | 4 (2.3%) | 3 (1.7%) |
| 22 years to 30 years | 57 | 0 | 1 (1.8%) |
| 31 years to 40 years | 71 | 1 (1.4%) | 3 (4.2%) |
| 41 years to 50 years | 52 | 0 | 1 (1.9%) |
| 51 years to 60 years | 46 | 1 (2.2%) | 3 (6.5%) |
| 61 years to 70 years | 33 | 1 (3.0%) | 1 (3.0%) |
| 71 years to 80 years | 16 | 1 (6.3%) | 4 (25.0%) |
| 81 years and above | 7 | 1 (14.3%) | 0 |
| Age Not Reported | 41 | 0 | 1 (2.4%) |
| Total | 1519 | 203 (13.4%) | 77 (5.1%) |
| * There were 2 - respiratory syncytial virus + metapneumovirus co- | | | |
| infections detected. | | | |

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

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

Sec05 FastPoint_RSV_MPV_09-10061 FDA.doc

2 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.

12

:

፡ ・

· , --- -

TABLE 5.6: Respiratory syncytial virus
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%

TABLE 5.7: Human.metapneumovirus
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
			95% CI
Sensitivity	55/80	68.8%	57.4-78.7%
Specificity	614/614	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: Respiratory syncytial virus
Fresh nasal/nasopharyngeal
swab	Comparator DSFA
(negatives followed by culture with DFA)
DHI DSFA	Positive	Negative	Total
Positive	39	0	39
Negative	1	647	648
Total	40	647	687
			95% CI
Sensitivity	39/40	97.5%	86.8-99.9%
Specificity	647/647	100.0%	99.4-100%

.

13

TABLE 5.9: Human metapneumovirus
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	632	652
Total	44	632	676
			95% CI
Sensitivity	24/44	54.5%	38.8-69.9%
Specificity	632/632	100.0%	99.4-100%

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

14

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 eagle with three stripes forming its body and wings. The eagle is facing right. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the eagle.

DEPARTMENT OF HEALTH & HUMAN SERVICES '

Public Health Service

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

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

DEC - 4 2009

Re: K093233

Trade/Device Name: D FastPoint L- DFA RSV/MPV Identification Kit Regulation Number: 21 CFR § 866.3980 Regulation Name: Respiratory viral antigens (respiratory syncytial virus and human metapneumovirus) Regulatory Class: II Product Code: OMG, LKT Dated: October 5, 2009 Received: October 14, 2009

Dear Mr. Lollar:

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

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

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

15

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

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

If you desire specific information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5460. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 443-6597 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html

Sincerely yours.

Ula Schla fu

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

16

510(k) Number (if known): K093233

Device Name: D3 FastPoint L-DFA RSV/MPV Identification Kit

Indication for Use:

The Diagnostic Hybrids, Inc. device, D FastPoint L-DFA RSV/MPV Identification Kit is intended for the qualitative identification of respiratory syncytial virus and human metapneumovirus 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 respiratory syncytial virus 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 syncytial virus and human metapneumovirus infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.

Prescription Use X (Part 21 CFR 801 Subpart D)

AND/OR

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

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

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

Uve Schif
Division Sign-Off

Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) k 093233