K Number

Device Name

SIMPLEXA FLU A/B & RSV DIRECT, SIMPLEXA FLU A/B & RSV POSITIVE CONTROL PACK

Manufacturer

FOCUS DIAGNOSTICS, INC.

Date Cleared

2012-07-13

(154 days)

Product Code

OCC OOI

Regulation Number

866.3980

Intended Use

The Focus Diagnostics Simplexa™ Flu A/B & RSV Direct assay is intended for use on the 3M Integrated Cycler instrument for the in vitro qualitative detection and differentiation of influenza A virus, influenza B virus, and respiratory syncytial virus (RSV) RNA in nasopharyngeal swabs (NPS) from human patients with signs and symptoms of respiratory tract infection in conjunction with clinical and epidemiological risk factors. This test is intended for use as an aid in the differential diagnosis of influenza A, influenza B, and RSV viral infections in humans and is not intended to detect influenza C. Negative results do not preclude influenza virus or RSV infection and should not be used as the sole basis for treatment or other patient management decisions. Performance characteristics for influenza A were established with clinical specimens collected during the 2010/2011 influenza season when 2009 H1N1 influenza and H3N2 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 the state or local health department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens. Focus Diagnostics' Simplexa™ Flu A/B & RSV Positive Control Pack is intended to be used as a control with the Simplexa™ Flu A/B & RSV Direct kit. This control is not intended for use with other assays or systems.

Device Description

The Simplexa™ Flu A/B & RSV Direct assay system is a real-time RT-PCR system that enables the direct amplification, delection and differentiation of human influenza A (Flu A) virus RNA, human influenza B (Flu B) virus RNA and RSV RNA from unprocessed nasopharyngeal swabs that have not undergone nucleic acid extraction. The system consists of the Simplexa™ Flu A/B & RSV Direct assay, the 3M Integrated Cycler (with Integrated Cycler Studio Software), the Direct Amplification Disc and associated accessories. In the Simplexa™ Flu A/B & RSV Direct assay, bi-functional fluorescent probe-primers are used together with corresponding reverse primers to amplify Flu A, Flu B, RSV and internal control RNA. The assay provides three results; conserved regions of influenza A viruses (matrix gene) influenza B viruses (matrix gene) and RSV (M gene) are targeted to identify these viruses in the specimen. An RNA internal control is used to detect RT-PCR failure and/or inhibition. The 3M Integrated Cycler is a rapid real-time Polymerase Chain Reaction thermocycler used for the identification of nucleic acid from prepared biological samples. The instrument utilizes disk media to contain and to process samples. The instrument uses real time flourometric detection to identify targets within the sample wells. The instrument is controlled by an external computer running the Integrated Cycler Studio software.

More Information

Contact

Type

Center

Panel

Date Received

Product Code

Subsequent Product Codes

Applicant Name (Manufacturer)

Device Name

Decision

Street 1

Street 2

City

State

Country Code

ZIP

Postal Code

Class Advise Committee

SSP Indicator

Third Party Reviewed

Expedited Review

Predicate Devices

K092500, K081030, K073029

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description and performance studies focus on real-time RT-PCR technology and standard analytical methods, with no mention of AI or ML algorithms for data analysis or interpretation.

Therapeutic

No
Explanation: This device is for qualitative detection and differentiation of viruses, aiding in diagnosis, not for treating or preventing disease.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the test "is intended for use as an aid in the differential diagnosis of influenza A, influenza B, and RSV viral infections in humans." This clearly indicates its role in diagnosis.

SaMD (Software as a Medical Device)

The device description explicitly states that the system consists of the assay, the 3M Integrated Cycler instrument, the Direct Amplification Disc, and associated accessories, in addition to the Integrated Cycler Studio Software. This indicates the device includes significant hardware components.

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 assay is intended for "in vitro qualitative detection and differentiation of influenza A virus, influenza B virus, and respiratory syncytial virus (RSV) RNA in nasopharyngeal swabs (NPS) from human patients". The term "in vitro" is a key indicator of an IVD, meaning it is used to test samples outside of the living body.
Device Description: The description details a "real-time RT-PCR system" that analyzes "human influenza A (Flu A) virus RNA, human influenza B (Flu B) virus RNA and RSV RNA from unprocessed nasopharyngeal swabs". This describes a laboratory test performed on a biological sample.
Performance Studies: The document includes detailed performance studies (Prospective Clinical Study, Retrospective Clinical Study, Analytical Sensitivity, etc.) which are standard requirements for demonstrating the accuracy and reliability of an IVD.
Predicate Device(s): The mention of "Predicate Device(s)" with K numbers (K092500, K081030, K073029) and names (Prodesse, ProFlu+ Assay) indicates that this device is being compared to previously cleared IVDs, a common process in regulatory submissions for IVDs.

All of these elements strongly indicate that the Focus Diagnostics Simplexa™ Flu A/B & RSV Direct assay is an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

Negative results do not preclude influenza virus or RSV infection and should not be used as the sole basis for treatment or other patient management decisions.

Performance characteristics for influenza A were established with clinical specimens collected during the 2010/2011 influenza season when 2009 H1N1 influenza and H3N2 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 the state or local health department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

Focus Diagnostics' Simplexa™ Flu A/B & RSV Positive Control Pack is intended to be used as a control with the Simplexa™ Flu A/B & RSV Direct kit. This control is not intended for use with other assays or systems.

Product codes (comma separated list FDA assigned to the subject device)

OCC, OOI

Device Description

In the Simplexa™ Flu A/B & RSV Direct assay, bi-functional fluorescent probe-primers are used together with corresponding reverse primers to amplify Flu A, Flu B, RSV and internal control RNA. The assay provides three results; conserved regions of influenza A viruses (matrix gene) influenza B viruses (matrix gene) and RSV (M gene) are targeted to identify these viruses in the specimen. An RNA internal control is used to detect RT-PCR failure and/or inhibition.

The 3M Integrated Cycler is a rapid real-time Polymerase Chain Reaction thermocycler used for the identification of nucleic acid from prepared biological samples. The instrument utilizes disk media to contain and to process samples. The instrument uses real time flourometric detection to identify targets within the sample wells. The instrument is controlled by an external computer running the Integrated Cycler Studio software.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

nasopharyngeal swabs (NPS)

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

Prospective Clinical Study:

Sample Size: A total of 722 nasopharyngeal swabs specimens were obtained. One (1) sample was excluded from the prospective analysis due to Invalid result for Flu A, three (3) samples were excluded due to an invalid result for Flu B and one (1) sample were excluded due to an invalid result for RSV.
Data Source: Specimens were collected from prospectively collected specimens with signs and symptoms of viral respiratory tract infection. Prospective samples were collected in Eastern United States from 10-November-2010 to 11-March-2011; the Mid-Western United States from 08-January-2011 and in Australia from 17-August-2010 to 20-October-2010.
Annotation Protocol: Reference results for influenza A, influenza B viruses and respiratory syncytial virus were generated using culture. Culture results were carried forward from the results obtained at the time of sample collection.

Retrospective Clinical Study:

Sample Size: A total of 223 nasopharyngeal swabs specimens were obtained.
Data Source: Specimens were obtained from retrospectively banked specimens from patients with signs and symptoms of viral respiratory tract infection.
Annotation Protocol: Reference results for influenza B viruses and respiratory syncytial virus were generated using culture. Culture results were carried forward from the results obtained at the time of sample banking.

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

CLINICAL AGREEMENT - PROSPECTIVE STUDY

Study Type: Prospective Clinical Study
Sample Size: 722 nasopharyngeal swabs specimens.
Key Results:
- Clinical Agreement - Flu A (All sites combined)
  - Sensitivity: 97.1%(66/68) (95% CI: 89.9 to 99.2%)
  - Specificity: 97.9%(639/653) (95% CI: 96.4 to 98.7%)
- Clinical Agreement - Flu B (All sites combined)
  - Sensitivity: 100.0%(21/21) (95% CI: 84.5 to 100.0%)
  - Specificity: 99.9%(697/698) (95% CI: 99.2 to 100.0%)
- Clinical Agreement - RSV (Site 1)
  - Sensitivity: 100.0%(1/1) (95% CI: 20.7 to 100.0%)
  - Specificity: 98.2%(323/329) (95% CI: 96.1 to 99.2%)
- Clinical Agreement - RSV (Site 2)
  - Sensitivity: 98.6%(72/73) (95% CI: 92.6 to 99.8%)
  - Specificity: 89.5%(154/172) (95% CI: 84.1 to 93.3%)
- Clinical Agreement - RSV (Site 3)
  - Sensitivity: 90.0%(9/10) (95% CI: 59.6 to 98.2%)
  - Specificity: 84.6%(115/136) (95% CI: 77.5 to 89.7%)

CLINICAL AGREEMENT - RETROSPECTIVE STUDY

Study Type: Retrospective Clinical Study
Sample Size: 223 nasopharyngeal swabs specimens.
Key Results:
- Clinical Agreement - Flu A (All sites combined)
  - PPA: 96.2% (76/79) (95% CI: 89.4 to 98.7%)
  - NPA: 99.3% (143/144) (95% CI: 96.2 to 99.9%)
- Clinical Agreement - Flu B (All sites combined)
  - PPA: 97.6% (40/41) (95% CI: 87.4 to 99.6%)
  - NPA: 100.0% (182/182) (95% CI: 97.9 to 100.0%)
- Clinical Agreement - RSV (All sites combined)
  - PPA: 100.0%(12/12) (95% CI: 75.7 to 100.0%)
  - NPA: 98.6%(208/211) (95% CI: 95.9 to 99.5%)

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION

Study Type: Analytical Sensitivity
Key Results: LoD ranges from 10^-2 - 10^-3 TCID50/mL.

ANALYTICAL REACTIVITY / CROSS REACTIVITY

Study Type: Analytical Reactivity and Cross Reactivity (Analytical Specificity)
Key Results: All strains tested were appropriately detected. No cross reactivity was detected for Flu A, Flu B or RSV.

INTERFERENCE

Study Type: Interference Study
Key Results: There was no evidence of interference caused by the substances tested.

INHIBITION BY OTHER MICROORGANISMS

Study Type: Inhibition Study
Key Results: No inhibitory effects were confirmed for influenza A, influenza B, or RSV at the concentrations tested.

CARRY-OVER CONTAMINATION

Study Type: Carry-over Contamination Study
Key Results: No cary-over contamination effect was seen in the Flu A, Flu B or RSV channels.

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

Sensitivity:
- Flu A (Prospective): 97.1%
- Flu B (Prospective): 100.0%
- RSV (Prospective Site 1): 100.0%
- RSV (Prospective Site 2): 98.6%
- RSV (Prospective Site 3): 90.0%
Specificity:
- Flu A (Prospective): 97.9%
- Flu B (Prospective): 99.9%
- RSV (Prospective Site 1): 98.2%
- RSV (Prospective Site 2): 89.5%
- RSV (Prospective Site 3): 84.6%
PPA (Positive Percent Agreement):
- Flu A (Retrospective): 96.2%
- Flu B (Retrospective): 97.6%
- RSV (Retrospective): 100.0%
NPA (Negative Percent Agreement):
- Flu A (Retrospective): 99.3%
- Flu B (Retrospective): 100.0%
- RSV (Retrospective): 98.6%
Limit of Detection (LoD): Ranges from 10^-2 - 10^-3 TCID50/mL for Influenza A, Influenza B, and RSV.
Reproducibility (%CV):
- Influenza A = 0.4 to 1.5
- Influenza B = 0.5 to 3.6
- RSV = 1.2 to 3.3

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

K092500, K081030, K073029

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

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

Image /page/0/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo features the word "FOCUS" in large, bold, sans-serif font. A curved, swooping line extends from the left side of the "F", arcing over the word. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin line.

510(k) Summary Simplexa™ Flu A/B & RSV Direct - REF MOL2650 implexa™ Flu A/B & RSV Positive Control Pack - REF MOL2660 Prepared Date: 12.July.2012 Page 1 of 7

| Applicant | Focus Diagnostics, Inc.
11331 Valley View Street
Cypress, California 90630
USA |
|--------------------------------|-----------------------------------------------------------------------------------------|
| Establishment Registration No. | 2023365 |
| | JUL 13 2012 |
| Contact Person | Tara Viviani
tel 562.240.6115
fax 562.240.6530
tviviani@focusdx.com |
| Summary Date | May 18, 2012 |
| Proprietary Name | Simplexa™ Flu A/B & RSV Direct
Simplexa™ Flu A/B & RSV Positive Control Pack |
| Generic Name | Respiratory Viral Panel |
| Classification | Class II, Special Controls |
| Regulation | § 21 CFR 866.3980 - respiratory viral panel multiplex nucleic acid
assay |
| Product Code | OCC - respiratory virus panel nucleic acid assay system |
| Predicate Devices | Prodesse, ProFlu+ Assay (K092500, K081030, K073029) |
| INTENDED USE | |

Negative results do not preclude influenza virus or RSV infection and should not be used as for treatment or other patient management decisions.

If infection with a novel Influenza A virus is suspected based on current clinical and enideria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to the state or local health department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

INTENDED USE

Device Description

Image /page/1/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo features the word "FOCUS" in bold, uppercase letters, with a curved graphic element to the left. Below "FOCUS" is the word "Diagnostics" in a smaller, non-bold font, underlined with a thin line.

510(k) Summary Simplexa™ Flu A/B & RSV Direct - REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack - REF MOL2660 Prepared Date: 12.July.2012 Page 2 of 7

K120413

are targeted to identify these viruses in the specimen. An RNA internal control is used to detect RT-PCR failure and/or inhibition.

Predicate Device Information

| Trade Name / Method | 510(k)
submitter | 510(k)
number | Decision Date | Panel | Product
Code(s) |
|---------------------|------------------------|---------------------------------|----------------------------------------|----------------------|--------------------|
| ProFlu+ | GenProbe
(Prodesse) | K092500,
K081030,
K073029 | 08/20/2009
05/02/2008
01/04/2008 | Microbiology
(83) | OCC |

Comparison to Predicate Device

	Device	Predicate
Item
Name	Simplexa™ Flu A/B & RSV Direct	ProFlu+
Intended Use	The Focus Diagnostics Simplexa™ Flu A/B & RSV Direct assay is intended for use on the 3M Integrated Cycler instrument for the in vitro qualitative detection and discrimination of influenza A virus, influenza B virus, and respiratory syncytial virus (RSV) RNA in nasopharyngeal swabs (NPS) from human patients with signs and symptoms of respiratory tract infection in conjunction with clinical and epidemiological risk factors. This test is intended for use as an aid in the differential diagnosis of influenza A, influenza B, and RSV viral infections in humans and is not intended to detect influenza C.
Negative results do not preclude influenza virus or RSV infection and should not be used as the sole basis for treatment or other patient management decisions.
Performance characteristics for influenza A were established with clinical specimens collected during the 2010/2011 influenza season when 2009 H1N1 influenza and H3N2 were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary.	The ProFlu™+ Assay is a multiplex Real-Time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and discrimination of Influenza A Virus, Influenza B Virus, and Respiratory Syncytial Virus (RSV) nucleic acids isolated and purified from nasopharyngeal (NP) swab specimens obtained from symptomatic patients. This test is intended for use to aid in the differential diagnosis of Influenza A, Influenza B and RSV viral infections in humans and is not intended to detect Influenza C.
Negative results do not preclude influenza or RSV virus infection and should not be used as the sole basis for treatment or other management decisions. It is recommended that negative RSV results be confirmed by culture. Performance characteristics for Influenza A Virus 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
	Device	Predicate
Item
Name	Simplexa™ Flu A/B & RSV Direct	ProFlu+
	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 department for testing. Viral
culture should not be attempted in
these cases unless a BSL 3+ facility is
available to receive and culture
specimens.	viruses and sent to state or local health
department for testing. Viral culture
should not be attempted in these cases
unless a BSL 3+ facility is available to
receive and culture specimens.
Assay Targets	Influenza A, Influenza B, RSV	Influenza A, Influenza B, RSV
Sample Types	NPS	NPS
Extraction
Methods	None	Roche MagNA Pure LC Total Nucleic
Acid Isolation Kit,
Biomérieux NucliSENS easyMAG
Assay
Methodology	PCR-based system for detecting the
presence / absence of viral RNA in
clinical specimens	PCR-based system for detecting the
presence / absence of viral DNA/RNA in
clinical specimens
Detection
Techniques	Multiplex assay using different reporter
dyes for each target.	Multiplex assay using different reporter
dyes for each target.
Influenza A Viral
Target	Well conserved region of the matrix
gene	Matrix gene
Influenza B Viral
Target	Well conserved region of the matrix
gene	Non-structural NS1 and NS2
Respiratory
Syncytial Viral
Target	M gene.	Polymerase
LoD	Analytical sensitivity (LoD) as defined
as the lowest concentration at which ≥
95% of all replicates tested positive,
ranges from 102 - 10-3 TCID50/mL.	Analytical sensitivity (LoD) as defined as
the lowest concentration at which ≥ 95%
of all replicates tested positive,
ranges from 102 - 10-1 TCID50/mL.
Reproducibility	Influenza A = %CV of 0.4 to 1.5
Influenza B = %CV of 0.5 to 3.6
RSV = %CV of 1.2 to 3.3	Influenza A = %CV of 1.4 to 5.3
Influenza B = %CV of 0.7 to 3.1
RSV = %CV of 1.5 to 8.3

Image /page/2/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo features a stylized swoosh shape on the left, followed by the word "FOCUS" in bold, sans-serif font. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font with a line above it.

K120413 510(k) Summary Simplexa™ Flu A/B & RSV Direct - REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack - REF MOL2660 Prepared Date: 12.July.2012

Page 3 of 7

PERFORMANCE CHARACTERISTICS

CLINICAL AGREEMENT - PROSPECTIVE STUDY

Three external testing sites and one internal site participated in a prospective clinical study. Reference results for influenza A, influenza B viruses and respiratory syncytial virus were generated using culture. Culture results were carried forward from the results obtained at the time of sample collection. A total of 722 nasopharyngeal swabs

1204'

510(k) Summary Simplexa™ Flu A/B & RSV Direct - REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack - REF MOL2660 Prepared Date: 12.July.2012 Page 4 of 7

specimens were obtained from prospectively collected specimens with signs and symptoms of viral respiratory tract infection. Prospective samples were collected in Eastern United States from 10-November-2010 to 11-March-2011; the Mid-Western United States from 08-January-2011 and in Australia from 17-August-2010 to 20-October-2010. Of the 722 specimens were collected from female patients and 397 specimens were collected from male patents. A total of 327 specimens were from patients 60 years of age, One (1) sample was excluded from the prospective analysis due to Invalid result for Flu A, three (3) samples were excluded due to an invalid result for Flu B and one (1) sample were excluded due to an invalid result for RSV. During the study the percentage of specimens with invalid results was 0.4% (3/722) with a 95% Cl of 0.1% to 1.2%.

Clinical Agreement - Flu A (Prospective - All sites combined)

Culture Result	Simplexa™ Results - Flu A		Sensitivity/Specificity
	n	Detected	Not Detected	95% CI
Detected	68	66	2	Sensitivity: 97.1%(66/68)
95% CI: 89.9 to 99.2%
Not Detected	653	14	639	Specificity: 97.9%(639/653)
95% CI: 96.4 to 98.7%

Clinical Agreement - Flu B (Prospective - All sites combined)

| Culture Result | N | Simplexa™ Results - FluB | | Sensitivity/Specificity
95% CI |
|----------------|-----|--------------------------|--------------|-------------------------------------------------------|
| | | Detected | Not Detected | |
| Detected | 21 | 21 | 0 | Sensitivity: 100.0%(21/21)
95% CI: 84.5 to 100.0% |
| Not Detected | 698 | 1 | 697 | Specificity: 99.9%(697/698)
95% CI: 99.2 to 100.0% |

Clinical Agreement - RSV (Prospective - Site 1)

Culture Result		Simplexa™ Results - RSV		Sensitivity/Specificity
		Detected	Not Detected	95% Cl ÷
Detected				Sensitivity: 100.0%(1/1)
95% Cl: 20.7 to 100.0%
Not Detected	329		323	Specificity: 98.2%(323/329)
95% CI: 96.1 to 99.2%

a) 4/6 samples were confirmed as RSV positive by an FDA cleared NAT

Clinical Agreement - RSV (Prospective - Site 2)

Culture Result	N	Simplexa™ Results – RSV		Sensitivity/Specificity
		Detected	Not Detected	95% CI
Detected	73	72	1a	Sensitivity: 98.6%(72/73)
95% CI: 92.6 to 99.8%
Not Detected	172	18b	154	Specificity: 89.5%(154/172)
95% CI: 84.1 to 93.3%

a) 1/1 sample confirmed as RSV positive by an FDA cleared DFA

b) 11/18 samples confirmed as RSV positive by an FDA cleared DFA

Image /page/4/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, sans-serif font, with the word "Diagnostics" in a smaller, non-bold font underneath. To the left of the word "FOCUS" is a black, curved shape that resembles a crescent or a stylized checkmark.

510(k) Summarv Simplexa™ Flu A/B & RSV Direct - REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack - REF MOL2660 Prepared Date: 12.July.2012 Page 5 of 7

Clinical Agreement - RSV (Prospective - Site 3)
Culture Result		Simplexa™ Results - RSV		Sensitivity/Specificity
	N	Detected	Not Detected	95% CI
Detected	10	9	1	Sensitivity: 90.0%(9/10)
95% CI: 59.6 to 98.2%
Not Detected	136	21a	115	Specificity: 84.6%(115/136)
95% CI: 77.5 to 89.7%

a) 20/21 samples confirmed as RSV positive by an FDA cleared NAT

CLINICAL AGREEMENT - RETROSPECTIVE STUDY

Three external testing sites participated in a retrospective clinical study. Reference results for influenza B viruses and respiratory syncytial virus were generated using culture. Culture results were carried forward from the results obtained at the time of sample banking. A total of 223 nasopharyngeal swabs specimens were obtained from retrospectively banked specimens from patients with signs and symptoms of viral respiratory tract infection.

Culture Result	n	Simplexa™ Results – Flu A		PPA/NPA*
		Detected	Not Detected	95% CI
Detected	79	76	3	PPA: 96.2% (76/79)
95% CI: 89.4 to 98.7%
Not Detected	144	1	143	NPA: 99.3% (143/144)
95% CI: 96.2 to 99.9%

Clinical Agreement - Flu A (Retrospective - All sites combined)

PPA = Positive Percent Agreement, NPA = Negative Percent Agreement

Clinical Agreement - Flu B (Retrospective - All sites combined)

Culture Result	N	Simplexa™ Results - Flu B		PPA/NPA*
		Detected	Not Detected	95% CI
Detected	41	40	1	PPA: 97.6% (40/41)
95% CI: 87.4 to 99.6%
Not Detected	182	0	182	NPA: 100.0% (182/182)
95% CI: 97.9 to 100.0%

PPA = Positive Percent Agreement, NPA = Negative Percent Agreement

Clinical Agreement - RSV (Retrospective - All sites combined)

Culture Result . "		Simplexa™ Results - RSV		PPA/NPA*
		Detected	Not Detected	95% Cl
Detected	12	17		PPA: 100.0%(12/12)
95% Cl: 75.7 to 100.0%
Not Detected	211		208	NPA: 98.6%(208/211)
95% CI: 95.9 to 99.5%

PPA = Positive Percent Agreement, NPA = Negative Percent Agreement

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION

The Limit of Detection (LoD) was determined for the Simplexa™ Flu AJB & RSV Direct assay using quantified stocks of influenza A, influenza B and RSV virus strains serially diluted in negative swab matrix. The lowest concentration with ≥95% detection (at least 19 out of 20 replicates) was determined to be the limit of detection for each assay,

Image /page/5/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo features the word "FOCUS" in bold, uppercase letters. A curved line extends from the left side of the letter "F", arching over the word. Below "FOCUS" is the word "Diagnostics" in a smaller, non-bold font, underlined with a thin line.

Simplexa™ Flu A/B & RSV Direct - REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack - REF MOL2660 Prepared Date: 12.Julv.2012 Page 6 of 7

Simplexa TM Flu A/B & RSV Direct Limit of Detection
Viral Strain	LoD (TCID50/mL)
Influenza A/PR/8/34 (H1N1)	0.005
Influenza A/Hong Kong/8/68 (H3N2)	10
Influenza A/Swine NY/02/2009 (H1N1)	0.1
Influenza B/Great Lakes/1739/54	2
Influenza B/Malaysia/2506/2004	20
RSV A2	1
RSV B CH93-18(18)	3

ANALYTICAL REACTIVITY / CROSS REACTIVITY

Analytical Reactivity

Different strains of influenza A including H1 and H3 subtypes, influenza B and RSV including A and B subtypes were evaluated. The most recent strains and geographically diverse strains were chosen. Quantified viral material was spiked into negative swab matrix at a sindle dilution with a concentration of approximately 1.0 x10 x10 x10 x10 TCIDsoml and assayed in triplicate. Ct values obtained during testing indicate all viral strains were tested near the LoD. All strains tested were appropriately detected.

Cross Reactivity (Analytical Specificity)

The Simplexa™ assay's analytical specificity was evaluated by testing the ability to exclusively identify influenza A virus and/or influenza B virus and/or RSV with no cross reactivity to organisms that are closely related, or cause similar clinical symptoms, or present as normal flora in the specimen types of interest.

The panel of thirty-two (32) potential cross reactants were into a swab matrix at clinically relevant concentrations. The unspiked matrix was also tested to serve as a baseline. Samples were tested in triplicate to screen for cross reactivity. If signal was detected in any detection channel (Flu A, Flu B, RSV) in any of the three replicates, an additional 5 replicates were tested for confirmation.

No cross reactivity was detected for Flu A, Flu B or RSV.

INTERFERENCE

The performance of this assay was evaluated with potentially interfering substances that may be present in nasopharyngeal swabs at the concentrations indicated in the table below. The potentially interfering substances were evaluated in a contrived sample that contained influenza A/PR/8/34 H1N1) at a concentration of 0.01 TCIDsomL and influenza B (influenza B/Malaysia/2506/2004) at a concentration of 40 TC/Ds/mL and RSV A2 at a concentration of 4 TCIDsomL. All strains were tested at two to four times the LoD. There was no evidence of interference caused by the substances tested.

| Potential Interferents | Active Ingredient | Interferent
Concentration |
|---------------------------------------|----------------------------------------|------------------------------|
| Afrin Nasal Spray | Oxymetazoline | 15% (v/v) |
| Antibacterial, systemic | Tobramycin | 4 µg/mL |
| Antibiotic, nasal ointment | Mupirocin | 6.6 mg/mL |
| Blood | N/A | 2%(v/v) |
| Purified Mucin Protein | Bovine Submaxillary Gland Type I-
S | 60 µg/mL |
| Nasal Corticosteroid - Beconase
AQ | Beclomethasone | 5% (v/v) |
| Nasal Corticosteroid - Fluticasone | Fluticasone | 5% (v/v) |
| Relenza Antiviral Drug | Zanamivir | 3.3 mg/mL |
| Tamiflu Antiviral Drug | Oseltamivir | 1 μΜ |

Image /page/6/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo features the word "FOCUS" in large, bold, sans-serif font, with a curved, black shape above and to the left of the word. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin line.

20413

510(k) Summary

Simplexa™ Flu A/B & RSV Direct - REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack - REF MOL2660 Prepared Date: 12.July.2012

Page 7 of 7

| Potential Interferents | Active Ingredient | Interferent
Concentration |
|------------------------|----------------------------------------------------------------------|------------------------------|
| Zicam Nasal Gel | Luffa Opperculata, Galphimia
glauca, histaminum
hydrochloricum | 5% (v/v) |

INHIBITION BY OTHER MICROORGANISMS

The Simplexa™ assay was evaluated by testing the ability to identify influenza B virus, and RSV when potentially inhibitory organisms are present.

The panel of thirty two (32) potentially inhibitory organisms was individually spiked into a low concentration (approximately 2 times LoD) of influenza A/PR/8/34 H1N1), influenza B (Influenza B/Malaysia/2506/2004) and RSV (A2). Samples were tested in triplicate to screen for inhibition. If signal was not detected in any detection channel (Flu A, Flu B, RSV) in any of the three replicates, an additional 5 replicates were tested for confirmation.

No inhibitory effects were confirmed for influenza A, influenza B, or RSV at the concentrations tested.

CARRY-OVER CONTAMINATION

An internal carry-over study searched for the presence of contamination in negative samples. The study was designed by alternately placing a high positive and a negative sample on each disc. The carryover effect was evaluated by comparing the observed negative rate for the negative sample with the expected rate under normal reproducibility conditions. No cary-over contamination effect was seen in the Flu A, Flu B or RSV channels.

Image /page/7/Picture/1 description: The image shows the logo for the U.S. Department of Health and 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 an abstract image of an eagle with its wings spread.

Food and Drug Administration

10903 New Hampshire Avenue Silver Spring, MD 20993

JUL 1 3 2012

Focus Diagnostics, Inc. c/o Tara Viviani Regulatory Affairs Project Manager 11331 Valley View Street Cypress, California 90630

Re: K120413

Trade/Device Name: Simplexa™Flu A/B & RSV Direct Simplexa "Flu A/B & RSV Positive Control Pack Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel nucleic acid assay system Regulatory Class: Class II Product Code: OCC, OOI Dated: May 18, 2012 Received: May 21, 2012

Dear Ms. Viviani:

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 class II (Special Controls), 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 or any is with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of

Page 2 - Tara Viviani

CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); 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 advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

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) 796-7100 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

Jóááym.

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

Indications for Use Statement

510(k) Number (if known): K120413

Simplexa™ Flu A/B & RSV Direct Device Name: Simplexa™ Flu A/B & RSV Positive Control Pack

Indications for Use:

Simplexa™ Flu A/B & RSV Direct (REF MOL2650)

Negative results do not preclude influenza virus or RSV infection and should not be used as the sole basis for treatment or other patient management decisions.

Performance characteristics for influenza A were established with clinical specimens collected during the 2010/2011 influenza season when 2009 HINI influenza and H3N2 were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary.

Simplexa™ Flu A/B & RSV Positive Control Pack (REF) MOL2660)

X And/Or Prescription Use (Part 21 CFR 801 Subpart D)

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

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

Concurrence of CDRH, Office of In Vitro Diagnostic Device Evaluation and Safety (OIVD)

Tamara Feldhuhn

Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K 120413