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 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 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.

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

Focus Diagnostics' Simplexa™ Flu A/B & RSV Positive Control Pack is intended to be used as a control with the SimplexaTM 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, detection 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.

AI/ML Overview

The provided text describes a 510(k) summary for the Simplexa™ Flu A/B & RSV Direct and Simplexa™ Flu A/B & RSV Positive Control Pack. This submission is intended to add eight additional influenza strains to the analytical reactivity of the device and addresses modifications made to the device from an earlier version (K120413).

Here's a breakdown of the requested information based on the document:

1. Table of acceptance criteria and the reported device performance

The document does not explicitly state "acceptance criteria" in the format of pass/fail thresholds for clinical performance. Instead, it presents "Positive Percent Agreement (PPA)" and "Negative Percent Agreement (NPA)" with a predicate device (Gen 1.0) and between device versions (Gen 2.0 and Gen 2.1) as part of method comparison studies, along with analytical reactivity and specificity data.

Method Comparison Results (Performance in relation to K120413 and between versions)

Target	Comparison	Metric	Reported Device Performance (Gen 2.0 vs Gen 1.0)	95% CI (Gen 2.0 vs Gen 1.0)	Reported Device Performance (Gen 2.1 vs Gen 2.0)	95% CI (Gen 2.1 vs Gen 2.0)
Flu A	PPA	Gen 2.0 vs Gen 1.0	100.0% (58/58)	93.0% to 100.0%	-	-
	NPA	Gen 2.0 vs Gen 1.0	95.7% (198/207)	91.9% to 97.7%	-	-
	PPA	Gen 2.1 vs Gen 2.0	-	-	100.0% (58/58)	93.8% to 100.0%
	NPA	Gen 2.1 vs Gen 2.0	-	-	99.0% (205/207)	96.5% to 99.7%
Flu B	PPA	Gen 2.0 vs Gen 1.0	98.2% (54/55)	90.4% to 99.7%	-	-
	NPA	Gen 2.0 vs Gen 1.0	95.7% (201/210)	92.1% to 97.7%	-	-
	PPA	Gen 2.1 vs Gen 2.0	-	-	100.0% (56/56)	93.6% to 100.0%
	NPA	Gen 2.1 vs Gen 2.0	-	-	100.0% (209/209)	98.2% to 100.0%
RSV	PPA	Gen 2.0 vs Gen 1.0	97.8% (45/46)	88.7% to 99.6%	-	-
	NPA	Gen 2.0 vs Gen 1.0	95.9% (210/219)	92.4% to 97.8%	-	-
	PPA	Gen 2.1 vs Gen 2.0	-	-	100.0% (55/55)	93.5% to 100.0%
	NPA	Gen 2.1 vs Gen 2.0	-	-	100.0% (210/210)	98.2% to 100.0%

Analytical Reactivity (Gen 2.1): All tested influenza A, influenza B, and RSV strains at specified concentrations were detected (100% detection for all, assayed in triplicate). These include:

18 Influenza A strains (H1, H3, H7N9)
10 Influenza B strains
4 RSV strains (A and B)

Cross Reactivity (Analytical Specificity) (Gen 2.1): No cross-reactivity was observed with 32 tested organisms (bacteria and other viruses) at clinically relevant concentrations. All results showed 0% detection for Flu A, Flu B, and RSV, and 100% detection for the Internal Control.

Interference (Gen 2.1): No evidence of interference was observed from potentially interfering substances (e.g., nasal sprays, antiviral drugs, blood, mucin protein) tested in contrived samples. All showed 100% detection for Flu A, Flu B, RSV, and RNA IC.

Limit of Detection (LoD) (Gen 2.1): The LoD for various strains across Gen 1.0, Gen 2.0, and Gen 2.1 are provided (e.g., Influenza A/Hong Kong/8/68 (H3N2) Gen 2.1 LoD: 0.1 TCID50/mL). The criteria for LoD determination was ≥95.0% detection (at least 31/32 replicates).

Precision (Gen 2.1): High reproducibility (low %CV) was observed for Ct values across inter-day, inter-run, inter-lot, and intra-run/lot variations for low and moderate positive samples of Flu A, Flu B, and RSV, as well as positive and negative controls. Qualitatively, all expected positive samples were detected at 100%, and negative samples were not detected for the target analytes.

2. Sample size used for the test set and the data provenance

Sample Size (Method Comparison): For each comparison (Gen 1.0 vs Gen 2.0, and Gen 2.0 vs Gen 2.1), 265 archived clinical samples were used.
- Composition: 55 positive for influenza A, 55 positive for influenza B, 55 positive for RSV, and 100 negative for all tested viruses.
- Data Provenance: The samples were "archived clinical samples" in Universal Transport Medium (UTM) or Viral Transport Medium (VTM).
  - 131 of these samples for the Gen 1.0 vs Gen 2.0 comparison were originally tested in support of K120413. The remaining 134 included 33 from the 2010-2011 flu season and 101 from the 2013-2014 flu season.
  - For the Gen 2.0 vs Gen 2.1 comparison, 125 samples were from K120413 study. The remaining 140 included 48 from the 2010-2011 flu season, 9 from 2012-2013, and 83 from 2013-2014.
  - The country of origin is not specified, but the context implies data likely from the USA (given FDA submission). The data is retrospective as it uses archived samples.
Sample Size (Analytical Reactivity/Cross Reactivity/Interference):
- Analytical Reactivity: Each viral strain was assayed in triplicate.
- Cross Reactivity: Each organism was tested in triplicate (3 replicates). Baseline negative matrix was tested in five (5) replicates.
- Interference: Each interfering substance was tested in triplicate (3 replicates). Baseline was tested in 15 replicates.
- Limit of Detection: Initially, 4 concentrations per virus tested in triplicate. Confirmatory testing involved 32 replicates for the lowest concentration.
- Precision: Each sample panel member tested in duplicate for each Reaction Mix lot in each run, two runs per day for a total of three days, yielding at least 36 replicates per panel member (41 for one Flu B sample).

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts

The document does not explicitly state the use of "experts" to establish ground truth for the test set in the method comparison studies. The ground truth for these studies appears to be based on results from the predicate device (Simplexa™ Flu A/B & RSV Direct Gen 1.0) and/or other FDA cleared Nucleic Acid Tests (NATs) for discrepant samples. For analytical studies (reactivity, cross-reactivity, interference, LoD, precision), the ground truth is established by the known concentration/presence of the spiked organisms or substances.

4. Adjudication method for the test set

For the method comparison studies, discrepancies between the modified device (Gen 2.0 or Gen 2.1) and the predicate device (Gen 1.0 or Gen 2.0 respectively) were sometimes resolved using another FDA cleared NAT. For example, for Flu A discrepancies in the Gen 1.0 vs Gen 2.0 comparison, "7/9 discrepant (K120413 – Negative and K142365 – Positive) samples were positive for Flu A on another FDA cleared NAT." This suggests a form of adjudication using a third, independent, cleared method.

5. If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

This section is not applicable. The device described is an in vitro diagnostic (IVD) assay for the detection of viruses using real-time RT-PCR, not an AI-powered diagnostic imaging device involving human readers or interpretation of medical images. Therefore, MRMC studies and the concept of human reader improvement with AI assistance do not apply.

6. If a standalone (i.e., algorithm only without human-in-the-loop performance) was done

This concept is not directly applicable in the context of an IVD assay like the Simplexa™ Flu A/B & RSV Direct. The device is essentially a "standalone" algorithm/assay from the perspective of direct human interpretation providing a qualitative result (detected/not detected). The performance metrics (PPA, NPA, analytical reactivity, LoD, etc.) represent the standalone performance of the assay system. There is no human "in the loop" for interpreting the raw assay output; the instrument's software interprets the Ct values to provide a qualitative result.

7. The type of ground truth used (expert consensus, pathology, outcomes data, etc.)

Clinical Studies (Method Comparison): The ground truth for the clinical sample comparisons was based on:
- The performance of the predicate device (Simplexa™ Flu A/B & RSV Direct Gen 1.0) for direct comparison between versions.
- Other FDA cleared Nucleic Acid Tests (NATs) for resolving discrepant results between the device versions.
Analytical Studies (Reactivity, Cross-Reactivity, Interference, LoD, Precision): The ground truth was established by known spiked concentrations of characterized viral strains, bacterial organisms, or potentially interfering substances into negative matrix.

8. The sample size for the training set

The document does not explicitly mention a "training set" in the context of machine learning or AI models, as this is an IVD assay. The development and optimization ("changes to the reaction mix formulation and cycling conditions," "changes to the manufacturing process and materials") that led to Gen 2.0 and Gen 2.1 would have involved internal validation and optimization data, which could be considered analogous to training data in a broad sense for assay development. However, specific "training set sizes" are not provided.

9. How the ground truth for the training set was established

As described in point 8, a formal "training set" for an AI model is not applicable here. For the assay development and optimization, ground truth would have been established through controlled laboratory experiments using well-characterized viral isolates and defined concentrations, analogous to how ground truth for the analytical studies (reactivity, LoD) was established.

Summary

{0}------------------------------------------------

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized depiction of a human figure in profile, with three overlapping faces suggesting a sense of community and support. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the figure, emphasizing the organization's name and national affiliation. The logo is simple, clean, and uses a monochromatic color scheme.

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

Focus Diagnostics, Inc. Sharon Young. Senior Regulatory Affairs Specialist 11331 Valley View Street Cypress, CA 90630

December 5, 2014

Re: K142365 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 Multiplex Nucleic Acid Assay Regulatory Class: Class II Product Code: OCC, OOI Dated: September 26, 2014 Received: September 29, 2014

Dear Ms. Young:

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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. 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 CFR Part 807); labeling (21 CFR Part 801); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set

{1}------------------------------------------------

forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR 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 Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Uwe Scherf -S for

Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

{2}------------------------------------------------

Indications for Use

510(k) Number (if known) K142365

Device Name

Simplexa™ Flu A/B & RSV Direct and Simplexa™ Flu A/B & RSV Positive Control Pack

Indications for Use (Describe)

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.

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

Type of Use (Select one or both, as applicable)
Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services

Food and Drug Administration

Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

{3}------------------------------------------------

Image /page/3/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, black letters. To the left of the word is a curved black line that starts thin and gets thicker as it curves around the word. Below the word "FOCUS" is the word "Diagnostics" in a smaller, less bold font, with a horizontal line above it.

510(k) Summarv

Simplexa" Flu A/B & RSV Direct REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 1 of 14

Applicant	Focus Diagnostics, Inc.11331 Valley View StreetCypress, California 90630USA
Establishment Registration No.	2023365
Contact Person	Sharon Youngtel 562.240.6680fax 562.240.6529syoung@focusdx.com
Summary Date	December 2, 2014
Proprietary Name	Simplexa™ Flu A/B & RSV Direct and Simplexa™ Flu A/B & RSVPositive Control Pack
Generic Name	Respiratory viral panel nucleic acid
Classification	Class II
US Product Code	OCC - Respiratory Viral Panel Nucleic Acid Assay SystemOOI - Real Time Nucleic Acid Amplification System
Regulation Number	21 CFR § 866.3980
Predicate Device	K120413 Simplexa™ Flu A/B & RSV Direct and Simplexa™ Flu A/B& RSV Positive Control Pack

510(k) SUBMISSION PURPOSE

The purpose of the submission is to add eight (8) additional influenza strains to the Analytical Reactivity of the Simplexa™ Flu A/B & RSV Direct. The package insert was updated to include the additional strains which included the following six (6) additional strains of Influenza A and two (2) additional strains of Influenza A/Anhui/1/2013, Influenza A/California/12/2012 (H1N1) pdm09, Influenza A/Indiana/08/2011 (H3N2)v, Influenza A/Minnesota/11/2010 (H3N2)v, Influenza A/Ohio/02/2012 (H3N2), Influenza A/Texas/50/2012 (H3N2), Influenza B/Brisbane/60/2008 and Influenza B/Wisconsin/01/2010.

Changes to the Simplexa™ Flu A/B and RSV Direct K120413 (Gen 1) were made in two phases. The first phase of the modifications (Gen 2.0) included changes to the reaction mix formulation and cycling conditions: (a) change in the annealing temperature (decreased), (b) change in the (increased), (c) change in the enzyme used, and (d) change to the RSV cut-off Ct (decreased). Limited changes (Gen. 2.1) were also made to the manufacturing process and materials to increase the stability of the reaction mix and to minimize non-specific products.

INTENDED USE

Simplexa™ Flu A/B & RSV Direct REF MOL2650

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 B virus, and respiratory syncytial virus (RSV) RNA in nasopharyngeal swabs (NPS) from human patients 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 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.

{4}------------------------------------------------

Image /page/4/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, sans-serif font. To the left of the word "FOCUS" is a curved, black shape that resembles a stylized check mark. Below the word "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin black line.

510(k) Summary Simplexa" Flu A/B & RSV Direct REF MOL2650 Simplexa" Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 2 of 14

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

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

MATERIALS PROVIDED

The Focus Diagnostics Simplexa™ Flu A/B & RSV Direct kit contains sufficient reagents for 24 reactions. Upon receipt, store at -10 to -30 ℃ (do not use a frost-free freezer). Each vial contains sufficient material for one use. Use within 30 minutes of removing from the freezer.

Component Name	REF	EC SYMBOLON LABEL	AbbreviatedName	CapColor	Number ofVials	Reactionsper Vial/Kit	Volume perVial
Simplexa™ Flu A/B &RSV Direct Reaction Mix	MOL2651	REAGA	RM	Brown	24	1/24	50 μL

Component Description

Kit Component	Contents
	DNA polymerase, Reverse Transcriptase, RNase inhibitor, buffer and dNTPs, encapsulated RNA TemplateDye-labeled fluorescent primers specific for detection of influenza A, influenza B and RSV and for theInternal Control
	Target ProbeFluorophore(Dye) Excitation Emission Targeted Gene Flu A FAM 495 520 matrix Flu B JOE 520 548 matrix RSV CFR610 590 610 M gene Internal Control"RNA IC" Q670 644 670 N/A
Simplexa™ Flu A/B &RSV Direct ReactionMix (RM)


Simplexa™ Flu A/B &RSV Direct BarcodeCard	Assay specific parameters, lot number, expiration date

Kit Description

{5}------------------------------------------------

Image /page/5/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 font size underneath. To the left of the word "FOCUS" is a curved, black shape that resembles a check mark or a swoosh.

42365 510(k) Summary Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa"" Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014

Page 3 of 14

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

Product Description

Component Name	REF	Description	Cap Color	Number of Vials	Reactions per Vial/Kit	Volume per Vial
Simplexa™ Flu A/B & RSVDirect Positive Control	MOL2661	Inactivated influenza A virus,Inactivated influenza B virus,Inactivated RSV	Red	10	1/10	100 µL

MATERIALS SUPPLIED SEPARATELY

Direct Amplification Disc Kit (REF MOL1455)

Direct Amplification Discs for use on the Integrated Cycler. a.

INSTRUMENT REQUIREMENTS

3M Integrated Cycler with Integrated Cycler Studio Software version 4.2 or higher

COMPARISON TO PREDICATE

Similarities

Feature	Predicate K120413	Modified Device K142365
Intended Use	Simplexa™ Flu A/B & RSV Direct REFMOL2650The 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	Same
Feature	Predicate K120413	Modified Device K142365
	Influenza viruses and sent to the state or localhealth department for testing. Viral cultureshould not be attempted in these cases unlessa BSL 3+ facility is available to receive andculture specimens.
	Simplexa™ Flu A/B & RSV Positive ControlPack REF MOL2660
	Focus Diagnostics' Simplexa™ Flu A/B & RSVPositive Control Pack is intended to be used asa control with the Simplexa™ Flu A/B & RSVDirect kit. This control is not intended for usewith other assays or systems.
Technology	The Simplexa™ Flu A/B & RSV Direct assaysystem is a real-time RT-PCR system thatenables the direct amplification, detection anddifferentiation of human influenza A (Flu A)virus RNA, human influenza B (Flu B) virusRNA and RSV RNA from unprocessednasopharyngeal swabs that have notundergone nucleic acid extraction. The systemconsists of the Simplexa™ Flu A/B & RSVDirect assay, the 3M Integrated Cycler (withIntegrated Cycler Studio Software), the DirectAmplification Disc and associated accessories.In the Simplexa™ Flu A/B & RSV Direct assay,bi-functional fluorescent probe-primers are usedtogether with corresponding reverse primers toamplify Flu A, Flu B, RSV and internal controlRNA. The assay provides three results;conserved regions of influenza A viruses (matrixgene), influenza B viruses (matrix gene) andRSV (M gene) are targeted to identify theseviruses in the specimen. An RNA internalcontrol is used to detect RT-PCR failure and/orinhibition.	Same
Instrument	3M Integrated Cycler	Same
SpecimenType	Unprocessed nasopharyngeal swabs that havenot undergone nucleic acid extraction	Same
Influenza AViral Target	Well conserved region of the matrix gene	Same
Influenza BViral Target	Well conserved region of the matrix gene	Same
RespiratorySyncytialViral Target	M gene	Same
Assay Type	Qualitative	Same

{6}------------------------------------------------

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

K142365

510(k) Summary

Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa"" Flu A/B & RSV Positive Control Pack [REE] MOL2660 Prepared Date: December 2, 2014 Page 4 of 14

{7}------------------------------------------------

Image /page/7/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, black letters. There is a curved black shape to the left of the word. Below the word "FOCUS" is the word "Diagnostics" in a smaller font, underlined with a thin black line.

K142365

510(k) Summary

Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack [REE]MOL2660 Prepared Date: December 2, 2014 Page 5 of 14

Differences

{8}------------------------------------------------

Image /page/8/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in a bold, sans-serif font, with the word "Diagnostics" in a smaller font underneath. There is a curved, swooping shape to the left of the word "FOCUS", which adds a dynamic element to the logo. The logo is in black and white.

510(k) Summarv

Simplexa" Flu A/B & RSV Direct REF MOL2650 Simplexa" Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 6 of 14

	Predicate K120413	Modified Device K142365
Manufacturing, Material andCycling Parameters		(Gen 2.0) - Changes to the annealing temperature (decreased), change in the run time (increased) and changes in enzyme used and change to the RSV cut-off Ct (decreased).
		(Gen 2.1) - Changes to manufacturing process and materials.

Method Comparison - Simplexa™ Flu AB & RSV Direct Gen 1.0 (K120413) to Simplexa™ Flu AJB & RSV Direct Gen 2.0

The changes implemented for Simplexa™ Flu A/B & RSV Direct (Gen 2.0) were evaluated with archived dinical samples. Based on historical data, 265 samples in Universal Transport Medium (UTM) or Viral Transport Medium (VTM) were assembled with 55 samples positive for influenza A, 55 samples positive for influenza B, 55 samples positive for RSV and 100 samples negative for all of the viruses tested. The 265 samples tested included 131 archived clinical samples originally tested in the clinical study conducted in support of the 510(k) submission (K120413). The other 134 samples included 33 archived samples from the 2010-2011 flu season and 101 samples from the 2013-2014 flu season. The samples were tested in parallel using the Simplexa™ Flu A/B & RSV Direct (Gen 1.0) and the Simplexa™ Flu A/B & RSV Direct (Gen 2.0). The positive percent agreement (PPA) and the negative percent agreement (NPA) between the results obtained with the Simplexa™ Flu A/B & RSV Direct (Gen 1.0) and the Simplexa™ Flu A/B & RSV Direct (Gen 2.0) for each virus are shown below:

Method Comparison - Simplexa™ Flu A/B & RSV Direct Gen 2.0 to Simplexa™ Flu A/B & RSV Direct Gen 2.1(K142365)

The changes implemented for Simplexa™ Flu A/B & RSV Direct (Gen 2.1) were evaluated with archived clinical samples. Based on historical data, 265 samples in Universal Transport Medium (UTM) or Viral Transport Medium (VTM) were assembled with 55 samples positive for influenza A, 55 samples positive for influenza B, 55 samples positive for RSV and 100 samples negative for all of the viruses tested. The 265 samples tested included 125 archived clinical samples originally tested in the clinical study conducted in support of the 510(k) submission (K120413). The other 140 samples included 48 archived samples from the 2010-2011 flu season. 9 samples from 2012-2013 flu season and 83 samples from the 2013-2014 flu season. The samples were tested in parallel using the Simplexa™ Flu A/B & RSV Direct (Gen 2.0) and the Simplexa™ Flu A/B & RSV Direct (Gen 2.1). The positive percent agreement (PPA) and the negative percent agreement (NPA) between the results obtained with the Simplexa™ Flu A/B & RSV Direct (Gen 2.0) and the Simplexa™ Flu A/B & RSV Direct (Gen 2.1) for each virus are shown below:

Simplexa™Flu A/B &RSV Direct(Gen 2.0)FluAResult	Simplexa™ Flu A/B & RSVDirect (Gen 1.0) Flu AResult		GrandTotal		%Agreement	95% CI
	Detected	NotDetected
Detected	58	9a	67	PPA	100.0% (58/58)	93.0% to 100.0%
NotDetected	0	198	198	NPA	95.7% (198/207)	91.9% to 97.7%
Grand Total	58	207	265
a7/9 discrepant (K120413 – Negative and K142365 – Positive) samples were positive for Flu A on another FDAcleared NAT.

Influenza A - Simplexa™ Flu A/B & RSV Direct Gen 1.0 (K120413) to Simplexa™ Flu A/B & RSV Direct Gen 2.0

PPA: Positive Percent Agreement, NPA: Negative Percent Agreement

{9}------------------------------------------------

Image /page/9/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in a bold, sans-serif font, with a curved, swooping shape extending from the top left of the "F" and arching over the word. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin line. The logo is in black and white.

510(k) Summary

Simplexa™ Flu A/B & RSV Direct REF M Simplexa" Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 7 of 14

Influenza A - Simplexa™ Flu A/B & RSV Direct Gen 2.0 to Simplexa™ Flu A/B & RSV Direct Gen 2.1 (K142365)

Simplexa™Flu A/B &RSV Direct(Gen 2.1)Flu AResult	Simplexa™ Flu A/B & RSVDirect (Gen 2.0) Flu AResult		GrandTotal		%Agreement	95% CI
	Detected	NotDetected
Detected	58	2	60	PPA	100.0% (58/58)	93.8% to 100.0%
NotDetected	0	205	205	NPA	99.0% (205/207)	96.5% to 99.7%
Grand Total	58	207	265

PPA: Positive Percent Agreement, NPA: Negative Percent Agreement

Influenza B - Simplexa™Flu A/B & RSV Direct Gen 1.0 (K120413) to Simplexa™ Flu A/B & RSV Direct Gen 2.0

Simplexa™Flu A/B &RSV Direct(Gen 2.0)Flu BResult	Simplexa™ Flu A/B & RSV Direct (Gen 1.0) Flu B Result		GrandTotal		%Agreement	95% CI
	Detected	NotDetected
Detected	54	9b	63	PPA	98.2% (54/55)	90.4% to 99.7%
NotDetected	1c	201	202	NPA	95.7% (201/210)	92.1% to 97.7%
Grand Total	55	210	265

95/9 discrepant (K120413 – Negative and K142365 – Positive) samples were positive for Flu B on another FDA cleared NAT.

°1/1 discrepant (K120413 – Positive and K142365 – Negative) samples was negative for Flu B on another FDA cleared NAT.

PPA: Positive Percent Agreement, NPA: Negative Percent Agreement

Influenza B - Simplexa™ Flu A/B & RSV Direct Gen 2.0 to Simplexa™ Flu A/B & RSV Direct Gen 2.1 (K142365)

Simplexa™Flu A/B &RSV Direct(Gen 2.1)Flu BResult	Simplexa™ Flu A/B & RSVDirect (Gen 2.0) Flu BResult		GrandTotal		%Agreement	95% CI
	Detected	NotDetected
Detected	56	0	63	PPA	100.0% (56/56)	93.6% to 100.0%
NotDetected	0	209	202	NPA	100.0%(209/209)	98.2% to 100.0%
Grand Total	56	209	265

PPA: Positive Percent Agreement, NPA: Negative Percent Agreement

{10}------------------------------------------------

Image /page/10/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, sans-serif font. A curved, swooping line is to the left of the word. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin line.

0(k) Su

Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 8 of 14

RSV - Simplexa™ Flu A/B & RSV Direct Gen 1.0 (K120413) to Simplexa™ Flu A/B & RSV Direct Gen 2.0

Simplexa™Flu A/B &RSV Direct(Gen 2.0)RSV Result	Simplexa™ Flu A/B & RSVDirect (Gen 1.0) RSV Result		GrandTotal		%Agreement	95% CI
	Detected	Not Detected
Detected	45	9d	54	PPA	97.8% (45/46)	88.7% to 99.6%
NotDetected	1e	210	211	NPA	95.9% (210/219)	92.4% to 97.8%
Grand Total	46	219	265
d3/9 discrepant (K120413 – Negative and K142365 – Positive) samples were positive for RSV on another FDAcleared NAT.
e1/1 discrepant (K120413 – Positive and K142365 – Negative) sample was negative for RSV on another FDAcleared NAT.

PPA: Positive Percent Agreement, NPA: Negative Percent Agreement

RSV - Simplexa™ Flu A/B & RSV Direct Gen 2.0 to Simplexa™ Flu A/B & RSV Direct Gen 2.1 (K142365)

Simplexa™Flu A/B &RSV Direct(Gen 2.1)RSV Result	Simplexa™ Flu A/B & RSVDirect (Gen 2.0) RSV Result		GrandTotal		%Agreement	95% CI
	Detected	Not Detected
Detected	55	0	55	PPA	100.0% (55/55)	93.5% to 100.0%
Not Detected	0	210	210	NPA	100.0% (210/210)	98.2% to 100.0%
Grand Total	55	210	265

PPA: Positive Percent Agreement, NPA: Negative Percent Agreement

ANALYTICAL REACTIVITY/ CROSS REACTIVITY

Analytical Reactivity - Simplexa™ Flu A/B & RSV Direct (Gen 2.1)

Different strains of influenza A including H1, H3 and H5 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 single dilution at the concentrations below. Each was assayed in triplicate. Ct values obtained during testing indicate all viral strains were the LoD. All strains tested were appropriately detected.

Organism	Concentration Tested	Result
Influenza A/Taiwan/42/06 H1N1	100 TCID50/mL	Flu A Detected
Influenza A/Anhui/1/2013 (H7N9)	25,000 EID50/mL	Flu A Detected
Influenza A/Brisbane/10/07 H3	100 TCID50/mL	Flu A Detected
Influenza A/Brisbane/59/07 H1	100 TCID50/mL	Flu A Detected
Organism	Concentration Tested	Result
Influenza A/California/12/2012 (H1N1) pdm09	100 TCID50/mL	Flu A Detected
Influenza A/California/7/2009 NYMC x-179-A	100 IU/mL	Flu A Detected
Influenza A/Indiana/08/2011 (H3N2)v	100 TCID50/mL	Flu A Detected
Influenza A/Minnesota/11/2010 (H3N2)v	100 CEID50/mL	Flu A Detected
Influenza A/New Caledonia/20/99 H1N1	100 TCID50/mL	Flu A Detected
Influenza A/Ohio/02/2012 (H3N2)	200 CEID50/mL	Flu A Detected
Influenza A/Port Chalmers/1/73 H3N2	100 TCID50/mL	Flu A Detected
Influenza A/PR/8/34 H1N1	100 TCID50/mL	Flu A Detected
Influenza A/Solomon Island/03/06	100 TCID50/mL	Flu A Detected
Influenza A/Swine NY/02/2009 H1N1	100 TCID50/mL	Flu A Detected
Influenza A/Swine/1976/31 H1N1	100 TCID50/mL	Flu A Detected
Influenza A/Swine/lowa/15/30 H1N1	100 TCID50/mL	Flu A Detected
Influenza A/Texas/50/2012 (H3N2)	100 TCID50/mL	Flu A Detected
Influenza A/Wisconsin/67/05 H3	100 TCID50/mL	Flu A Detected
Influenza A/WS/33 H1N1	100 TCID50/mL	Flu A Detected
Influenza B/Allen/45	100 TCID50/mL	Flu B Detected
Influenza B/Brisbane/60/2008	100 CEID50/mL	Flu B Detected
Influenza B/Florida/02/2006	100 TCID50/mL	Flu B Detected
Influenza B/Florida/04/2006	100 TCID50/mL	Flu B Detected
Influenza B/Florida/07/04	100 TCID50/mL	Flu B Detected
Influenza B/Hong Kong/5/72	100 TCID50/mL	Flu B Detected
Influenza B/Lee/40	100 TCID50/mL	Flu B Detected
Influenza B/Maryland/1/59	100 TCID50/mL	Flu B Detected
Influenza B/Panama/45/90	100 TCID50/mL	Flu B Detected
Influenza B/Taiwan/2/62	100 TCID50/mL	Flu B Detected
Influenza B/Wisconsin/01/2010	100 CEID50/mL	Flu B Detected
RSV-A Long	100 TCID50/mL	RSV Detected
RSV-B 9320	100 TCID50/mL	RSV Detected
Organism	Concentration Tested	Result
RSV-B Wash/18537/62	100 TCID50/mL	RSV Detected
RSV B WV/14617/85	100 TCID50/mL	RSV Detected

{11}------------------------------------------------

Image /page/11/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, black letters. To the left of the word is a black curved shape. Below the word "FOCUS" is the word "Diagnostics" in a smaller font with a line above it.

K142365

510(k) Summary

Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa"" Flu A/B & RSV Positive Control Pack [REE] MOL2660 Prepared Date: December 2, 2014 Page 9 of 14

{12}------------------------------------------------

Image /page/12/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 font size underneath. A curved, black shape is positioned to the left of the word "FOCUS", resembling a stylized check mark or swoosh. A horizontal line is placed above the word "Diagnostics".

510(k) Summary

Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack |REF MOL2660 Prepared Date: December 2, 2014 Page 10 of 14

NOTE: Although this test has been shown to detect novel avian influenza A (H7N9) and H3N2v cultured viruses, the performance characteristics of this device with clinical specimens that are positive for novel avian influenza A (H7N9) and H3N2v influenza viruses have not been established.

Cross Reactivity (Analytical Specificity) - Simplexa™ Flu A/B & RSV Direct (Gen 2.1)

Thirty-two (32) organisms were tested at clinically relevant concentrations to evaluate cross-reactivity of the Simplexa™ Flu A/B & RSV Direct. Three (3) instruments were used and fifteen (15) experimental runs across two (2) days by a single (1) operator were performed. Each of the organisms was spiked into a negative matrix and tested in triplicate (3). Baseline negative matrix was tested in five (5) replicates. No cross reactivity was observed.

	Tested		%Detection (# Detected / # Total)
Organism	Conc.	N	Flu A (FAM)	Flu B (JOE)	RSV (CFR610)	IC (Q670)
Baseline	N/A	5	0.0%(0/5)	0.0%(0/5)	0.0%(0/5)	100.0%(5/5)
Adenovirus 1	4.17E+05TCID50/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Adenovirus 7A	5.37E+05TCID50/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Bordetella pertussisA639	1.88E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Chlamydiapneumoniae	1.00E+06IFU/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Cytomegalovirus(CMV)	1.04E+05U/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Coronavirus 229E	5.89E+05TCID50/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Coronavirus OC43	1.95E+05TCID50/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Corynebacteriumdiphtheriae	4.00E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Enterovirus Type 71	1.10E+05TCID50/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Epstein-Barr Virus(EBV)	1.10E+05copies/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Escherichia coli0157:H7	1.10E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Haemophilusinfluenzae	1.41E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Lactobacillusplantarum, 17-5	7.97E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Legionellalongbeachae	8.63E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Measles	1.95E+05TCID50/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Metapneumovirus 9	1.58E+05TCID50/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Moraxella catarrhalis,NE1	1.49E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)

{13}------------------------------------------------

Image /page/13/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, sans-serif font. Above and to the left of the word "FOCUS" is a curved, swooping shape that resembles a check mark. Below the word "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 Simplexa™ Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 11 of 14

		N	%Detection (# Detected / # Total)
Organism	TestedConc.		Flu A (FAM)	Flu B (JOE)	RSV (CFR610)	IC (Q670)
Mumps	8.51E+05TCID50/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Mycobacteriumtuberculosis (genomicDNA)	6.54E+06c/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Mycoplasmapneumoniae, M129	3.16E+06ccu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Neisseria elongata	2.05E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Neisseriameningitides	7.07E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Parainfluenza 1	1.15E+05TCID50/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Parainfluenza 2	3.80E+05IU/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Parainfluenza 3	1.95E+05TCID50/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Pseudomonasaeruginosa	3.93E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Rhinovirus 1A	1.26E+05TCID50/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Staphylococcusaureus, COL	1.42E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Staphylococcusepidermidis	9.23E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Streptococcuspneumoniae	9.20E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Streptococcuspyogenes, M1	1.36E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
Streptococcussalivarius	2.12E+06cfu/mL	3	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)

INTERFERENCE - Simplexa™ Flu A/B & RSV Direct (Gen 2.1)

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 B and RSV. All strains were tested at two to four times the LoD. There was no evidence of interference caused by the substances tested.

Substance	Concentration Tested	Qualitative Result for each Channel
		Flu A (FAM)	Flu B (Joe)	RSV (CFR610)	RNA IC (Q670)
Baseline	None	100.0%(15/15)	100.0%(15/15)	100.0%(15/15)	100.0%(15/15)
Afrin Nasal Spray	15% (v/v)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)
Oseltamivir phosphate	1 $\mu$ M	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)
Blood	2% (v/v)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)
Zicam Nasal Gel	5% (v/v)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)
Tobramycin	4 $\mu$ g/mL	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)
Mupirocin	6.6 mg/mL	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)

{14}------------------------------------------------

Image /page/14/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, black letters. There is a curved black shape to the left of the word. Below the word "FOCUS" is the word "Diagnostics" in smaller, thinner, black letters with a line above it.

510(k) Summary

Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa" Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 12 of 14

Substance	ConcentrationTested	Qualitative Result for each Channel
		Flu A (FAM)	Flu B (Joe)	RSV (CFR610)	RNA IC (Q670)
Purified MucinProtein	60 µg/mL	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)
Beconase AQ	5% (v/v)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)
NasalCorticosteroid- Fluticasone	5% (v/v)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)
RelenzaAntiviral Drug- Zanamivir	3.3 mg/mL	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)
NTC	NA	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)	100.0%(3/3)
PositiveControl	NA	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)	100.0%(3/3)

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION

Three (3) strains of influenza A, two (2) strains of influenza B and two (2) strains of RSV were tested to determine the limit of detection (LoD) for the Simplexa™ Flu A/B & RSV Direct. Four concentrations per virus were tested in triplicate during the screening phase to determine the tentative LoD. The lowest concentration with all replicates detected during the screening phase was tested in thirty two (32) replicates to confirm detection at that concentration. The LoD is determined if at least 31/32 (≥95.0%) replicates are detected. The following table shows the comparison of the limit of detection between the original Simplexa™ Flu A/B & RSV Direct (Gen 1.0) and the modified Simplexa™ Flu A/B & RSV Direct (Gen 2.0 and Gen 2.1).

Virus	Gen 1.0(TCID50/mL)	Gen 2.0(TCID50/mL)	Gen 2.1(TCID50/mL)
Influenza A/ Hong Kong/8/68 (H3N2)	10	0.1	0.1
Influenza A/PR/8/34 (H1N1)	0.005	0.05	0.05
Influenza A/Swine NY/02/2009 (H1N1)	0.1	1	1
Influenza B/Great Lakes/1739/54	2	20	20
Influenza B/Malaysia/2506/2004	20	0.5	1
RSV-A2	1	2	2
RSV B CH93-18 (18)	3	2	2

{15}------------------------------------------------

Image /page/15/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, sans-serif font, with a curved, swooping shape above it. Below the word "FOCUS" is the word "Diagnostics" in a smaller, less bold font. The logo is in black and white.

510(k) Summary Simplexa" Flu A/B & RSV Direct REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 13 of 14

142365

PRECISION - Simplexa™ Flu A/B & RSV Direct (Gen 2.1)

The reaction mix (RM) lot to lot reproducibility was evaluated for Simplexa™ Flu A/B & RSV Direct (Gen 2.1) using three (3) Reaction Mix lots and sample panel which includes Low (~1.5 X LoD) and Moderate (~4 X LoD) positive samples for each of the three targets (Flu A, Flu B and RSV) and Positive Control. The low and moderate positive samples of each target were prepared by spiking the viral strain into negative swab matrix.

Each sample panel member was tested in duplicate (2) for each Reaction Mix lot in each run, two (2) runs per day for total of three (3) days using a single (1) 3M Integrated Cycler and a single (1) operator. The study produced at least thirty-six (36) replicates for each sample panel member. The Flu B moderate positive sample was tested in forty-one (41) replicates.

Quantitative Summary of Reaction Mix Inter-Lot Reproducibility
Analyte(Channel)	SamplePanelMember	N	MeanCt	Inter-Day		Inter-Run		Inter-Lot		Intra-Run/Lot		Total
				SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Flu A(FAM)	Flu A LowPositive	36	37	0	0	0.3	0.9	0	0	0.7	1.9	0.8	2.1
	Flu AModeratePositive	36	35.2	0	0	0	0	0.4	1	0.4	1.1	0.5	1.5
	PositiveControl(PC)	36	33.3	0.1	0.3	0.2	0.5	0.2	0.5	0.2	0.5	0.3	1
Flu B(JOE)	Flu B LowPositive	36	35.3	0	0	0	0	0.3	0.8	0.7	1.9	0.7	2.1
	Flu BModeratePositive	41	33.5	0	0	0.1	0.4	0.3	0.9	0.3	1	0.5	1.4
	PositiveControl(PC)	36	32.7	0	0	0	0	0.4	1.1	0.2	0.7	0.4	1.3
RSV(CFR610)	RSV LowPositive	36	36.5	0	0	0	0	0	0	0.8	2.3	0.8	2.3
	RSVModeratePositive	36	34.6	0	0	0.2	0.6	0	0	0.5	1.4	0.5	1.5
	PositiveControl(PC)	36	32.4	0.1	0.4	0.1	0.2	0	0.1	0.2	0.6	0.2	0.7
RNA IC(Q670)	Flu A LowPositive	36	29	0	0	0	0	0.4	1.5	0.4	1.2	0.6	1.9
	Flu AModeratePositive	36	29.1	0	0	0	0	0.4	1.4	0.4	1.2	0.6	1.9
	Flu B LowPositive	36	28.2	0.2	0.5	0	0	0.2	0.8	0.3	1	0.4	1.4
	Flu BModeratePositive	41	28.2	0	0.1	0	0.1	0.2	0.7	0.2	0.7	0.3	1
	RSV LowPositive	36	28.9	0	0	0	0	0.2	0.8	0.3	1.1	0.4	1.3
	RSVModeratePositive	36	28.9	0	0	0	0	0.3	0.9	0.3	0.9	0.4	1.2
	Negative	36	29.2	0	0	0.1	0.2	0	0	0.3	1	0.3	1
	PositiveControl	36	28.9	0	0.1	0	0	0.2	0.7	0.1	0.4	0.2	0.8

{16}------------------------------------------------

Image /page/16/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo features the word "FOCUS" in bold, sans-serif font, with the word "Diagnostics" in a smaller font size underneath. To the left of the word "FOCUS" is a curved, swooping graphic element that resembles an abstract eye or a stylized checkmark.

510(k) Summary

Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa"" Flu A/B & RSV Positive Control Pack |REF MOL2660 Prepared Date: December 2, 2014 Page 14 of 14

Analyte(Channel)	SamplePanelMember	N	Mean			Inter-Day			Inter-Run			Inter-Lot			Intra-Run/Lot			Total
	(PC)		Ct	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV

QualitativeResults(By Sample)	All	Flu A (FAM)		Flu B (JOE)		RSV (CFR610)
		DetectedN	Not DetectedN	DetectedN	Not DetectedN	DetectedN	Not DetectedN
		%	%	%	%	%	%
Flu A LowPositive	36	36			36		36
		100.0 %			100.0 %		100.0 %
Flu AMediumPositive	36	36			36		36
		100.0 %			100.0 %		100.0 %
Flu B LowPositive	36		36	36			36
			100.0 %	100.0 %			100.0 %
Flu BMediumPositive	41		41	41			41
			100.0 %	100.0 %			100.0 %
RSV: LowPositive	36		36		36	36
			100.0 %		100.0 %	100.0 %
RSV:MediumPositive	36		36		36	36
			100.0 %		100.0 %	100.0 %
NegativeTemplateControl(NTC)	36		36		36		36
			100.0 %		100.0 %		100.0 %
PositiveControl(PC)	36	36		36		36
		100.0 %		100.0 %		100.0 %

ADDITIONAL PERFORMANCE STUDIES

Please refer to the previously FDA cleared 510(k) K120413 for additional information.

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.