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K Number
K142365
Device Name
Simplexa Flu A/B & RSV Direct & Simplexa Flu A/B RSV Positive Control Pack
Manufacturer
FOCUS DIAGNOSTICS
Date Cleared
2014-12-05

(102 days)

Product Code
OCCOOI
Regulation Number
866.3980
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP Authorized
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 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.
More Information

K120413

K120413, K142365

No
The device description and performance studies focus on real-time RT-PCR technology and standard analytical methods for calculating agreement and performance metrics. There is no mention of AI or ML.

No
This device is an in vitro diagnostic (IVD) test, specifically a real-time RT-PCR system, used for the qualitative detection and differentiation of viral RNA to aid in diagnosis. It does not directly provide therapy or treatment.

Yes

This device is intended for the qualitative detection and differentiation of specific viral RNA in patient samples, serving as an aid in the differential diagnosis of influenza B and RSV viral infections. This directly aligns with the definition of a diagnostic device.

No

The device description explicitly states that the system consists of the assay, the 3M Integrated Cycler (with Integrated Cycler Studio Software), the Direct Amplification Disc, and associated accessories. This indicates the device includes significant hardware components beyond just software.

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 B virus, and respiratory syncytial virus (RSV) RNA in nasopharyngeal swabs...". The term "in vitro" means "in glass" or "in the lab," referring to tests performed outside of a living organism.
  • Device Description: The description details a "real-time RT-PCR system" that analyzes biological samples (nasopharyngeal swabs) to detect specific viral RNA. This is a typical characteristic of an in vitro diagnostic device.
  • Performance Studies: The document describes various performance studies (Method Comparison, Analytical Reactivity, Cross Reactivity, Interference, Analytical Sensitivity, Precision) conducted to evaluate the device's ability to accurately detect the target viruses in samples. These studies are required for IVD devices to demonstrate their performance characteristics.
  • Predicate Device: The mention of a "Predicate Device(s)" with a K number (K120413) indicates that this device is being compared to a previously cleared device, which is a standard process for submitting new IVD devices to regulatory bodies like the FDA.

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

N/A

Intended Use / Indications for 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 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.

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

OCC, OOI

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.

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

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

Method Comparison - Simplexa™ Flu AB & RSV Direct Gen 1.0 (K120413) to Simplexa™ Flu AJB & RSV Direct Gen 2.0:
Sample size: 265 samples
Data source: 131 archived clinical samples originally tested in the clinical study conducted in support of the 510(k) submission (K120413), 33 archived samples from the 2010-2011 flu season, and 101 samples from the 2013-2014 flu season.
Annotation protocol: Samples 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 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).

Method Comparison - Simplexa™ Flu A/B & RSV Direct Gen 2.0 to Simplexa™ Flu A/B & RSV Direct Gen 2.1(K142365):
Sample size: 265 samples
Data source: 125 archived clinical samples originally tested in the clinical study conducted in support of the 510(k) submission (K120413), 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.
Annotation protocol: Samples 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 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).

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

Method Comparison - Simplexa™ Flu AB & RSV Direct Gen 1.0 (K120413) to Simplexa™ Flu AJB & RSV Direct Gen 2.0
Study Type: Method Comparison Study
Sample Size: 265 samples
Sample Types: Archived clinical samples (131 from K120413 study, 33 from 2010-2011 flu season, 101 from 2013-2014 flu season) in Universal Transport Medium (UTM) or Viral Transport Medium (VTM).
Key Results:

  • Influenza A: PPA: 100.0% (58/58), NPA: 95.7% (198/207)
    • 7/9 discrepant (K120413 – Negative and K142365 – Positive) samples were positive for Flu A on another FDA cleared NAT.
  • Influenza B: PPA: 98.2% (54/55), NPA: 95.7% (201/210)
    • 9/9 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) sample was negative for Flu B on another FDA cleared NAT.
  • RSV: PPA: 97.8% (45/46), NPA: 95.9% (210/219)
    • 3/9 discrepant (K120413 – Negative and K142365 – Positive) samples were positive for RSV on another FDA cleared NAT.
    • 1/1 discrepant (K120413 – Positive and K142365 – Negative) sample was negative for RSV on another FDA cleared NAT.

Method Comparison - Simplexa™ Flu A/B & RSV Direct Gen 2.0 to Simplexa™ Flu A/B & RSV Direct Gen 2.1(K142365)
Study Type: Method Comparison Study
Sample Size: 265 samples
Sample Types: Archived clinical samples (125 from K120413 study, 48 from 2010-2011 flu season, 9 from 2012-2013 flu season, 83 from 2013-2014 flu season) in Universal Transport Medium (UTM) or Viral Transport Medium (VTM).
Key Results:

  • Influenza A: PPA: 100.0% (58/58), NPA: 99.0% (205/207)
  • Influenza B: PPA: 100.0% (56/56), NPA: 100.0% (209/209)
  • RSV: PPA: 100.0% (55/55), NPA: 100.0% (210/210)

Analytical Reactivity - Simplexa™ Flu A/B & RSV Direct (Gen 2.1)
Study Type: Analytical Reactivity
Key Results: All tested viral strains (various influenza A, B, and RSV subtypes) were appropriately detected at indicated concentrations (mostly 100 TCID50/mL).

Cross Reactivity (Analytical Specificity) - Simplexa™ Flu A/B & RSV Direct (Gen 2.1)
Study Type: Analytical Specificity
Sample Size: 3 replicates per organism, 5 replicates for baseline negative matrix (32 organisms tested)
Key Results: No cross reactivity was observed with 32 tested organisms at clinically relevant concentrations.

Interference - Simplexa™ Flu A/B & RSV Direct (Gen 2.1)
Study Type: Interference Study
Sample Size: 3 replicates per substance for each channel, 15 replicates for baseline (tested chemicals and drugs)
Key Results: No evidence of interference caused by the tested substances (Afrin Nasal Spray, Oseltamivir phosphate, Blood, Zicam Nasal Gel, Tobramycin, Mupirocin, Purified Mucin Protein, Beconase AQ, Nasal Corticosteroid - Fluticasone, Relenza Antiviral Drug - Zanamivir).

Analytical Sensitivity/Limit of Detection
Study Type: Limit of Detection (LoD) Study
Key Results: The LoD was determined for various influenza A, B, and RSV strains. The table provides a comparison of LoD values across Gen 1.0, Gen 2.0, and Gen 2.1.

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

Precision - Simplexa™ Flu A/B & RSV Direct (Gen 2.1)
Study Type: Reproducibility Study (Reaction Mix Lot to Lot)
Sample Size: At least 36 replicates for each sample panel member (41 for Flu B moderate positive sample)
Key Results: Coefficients of Variation (%CV) for Ct values were low across inter-day, inter-run, inter-lot, and intra-run/lot variability components, indicating good reproducibility. Qualitative results showed 100% detection for all positive samples and 100% not detected for negative control.

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

Key Metrics presented in the document are Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA).

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

  • Influenza A:
    • PPA: 100.0% (58/58)
    • NPA: 95.7% (198/207)
  • Influenza B:
    • PPA: 98.2% (54/55)
    • NPA: 95.7% (201/210)
  • RSV:
    • PPA: 97.8% (45/46)
    • NPA: 95.9% (210/219)

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

  • Influenza A:
    • PPA: 100.0% (58/58)
    • NPA: 99.0% (205/207)
  • Influenza B:
    • PPA: 100.0% (56/56)
    • NPA: 100.0% (209/209)
  • RSV:
    • PPA: 100.0% (55/55)
    • NPA: 100.0% (210/210)

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.

K120413 Simplexa™ Flu A/B & RSV Direct and Simplexa™ Flu A/B & RSV Positive Control Pack

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

§ 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.

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

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.

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)

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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 Street
Cypress, California 90630
USA |
|--------------------------------|--------------------------------------------------------------------------------------------------------------|
| Establishment Registration No. | 2023365 |
| Contact Person | Sharon Young
tel 562.240.6680
fax 562.240.6529
syoung@focusdx.com |
| Summary Date | December 2, 2014 |
| Proprietary Name | Simplexa™ Flu A/B & RSV Direct and Simplexa™ Flu A/B & RSV
Positive Control Pack |
| Generic Name | Respiratory viral panel nucleic acid |
| Classification | Class II |
| US Product Code | OCC - Respiratory Viral Panel Nucleic Acid Assay System
OOI - 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.

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.

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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

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 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, 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.

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 SYMBOL
ON LABEL | Abbreviated
Name | Cap
Color | Number of
Vials | Reactions
per Vial/Kit | Volume per
Vial |
|------------------------------------------------|---------|-----------------------|---------------------|--------------|--------------------|---------------------------|--------------------|
| Simplexa™ Flu A/B &
RSV Direct Reaction Mix | MOL2651 | REAG
A | RM | Brown | 24 | 1/24 | 50 μL |

Component Description

Kit ComponentContents
DNA polymerase, Reverse Transcriptase, RNase inhibitor, buffer and dNTPs, encapsulated RNA Template
Dye-labeled fluorescent primers specific for detection of influenza A, influenza B and RSV and for the
Internal Control
Target Probe
Fluorophore
(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 Reaction
Mix (RM)
Simplexa™ Flu A/B &
RSV Direct Barcode
CardAssay specific parameters, lot number, expiration date

Kit Description

5

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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 NameREFDescriptionCap ColorNumber of VialsReactions per Vial/KitVolume per Vial
Simplexa™ Flu A/B & RSV
Direct Positive ControlMOL2661Inactivated influenza A virus,
Inactivated influenza B virus,
Inactivated RSVRed101/10100 µL

MATERIALS SUPPLIED SEPARATELY

  1. 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

FeaturePredicate K120413Modified Device K142365
Intended UseSimplexa™ 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 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 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 Simplexa™ Flu A/B & RSV
Direct kit. This control is not intended for use
with other assays or systems. | |
| Technology | 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. | Same |
| Instrument | 3M Integrated Cycler | Same |
| Specimen
Type | Unprocessed nasopharyngeal swabs that have
not undergone nucleic acid extraction | Same |
| Influenza A
Viral Target | Well conserved region of the matrix gene | Same |
| Influenza B
Viral Target | Well conserved region of the matrix gene | Same |
| Respiratory
Syncytial
Viral Target | M gene | Same |
| Assay Type | Qualitative | Same |

6

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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

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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 K120413Modified Device K142365
Manufacturing, Material and
Cycling 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)
Flu
A
Result | Simplexa™ Flu A/B & RSV
Direct (Gen 1.0) Flu A
Result | | Grand
Total | | %Agreement | 95% CI |
|----------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------|-----------------|----------------|-----|-----------------|-----------------|
| | Detected | Not
Detected | | | | |
| Detected | 58 | 9a | 67 | PPA | 100.0% (58/58) | 93.0% to 100.0% |
| Not
Detected | 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 FDA
cleared 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 A
Result | Simplexa™ Flu A/B & RSV
Direct (Gen 2.0) Flu A
Result | | Grand
Total | | %Agreement | 95% CI |
|----------------------------------------------------------------------|-------------------------------------------------------------|-----------------|----------------|-----|-----------------|-----------------|
| | Detected | Not
Detected | | | | |
| Detected | 58 | 2 | 60 | PPA | 100.0% (58/58) | 93.8% to 100.0% |
| Not
Detected | 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 B
Result | Simplexa™ Flu A/B & RSV Direct (Gen 1.0) Flu B Result | | Grand
Total | | %Agreement | 95% CI |
|----------------------------------------------------------------------|-------------------------------------------------------|-----------------|----------------|-----|-----------------|----------------|
| | Detected | Not
Detected | | | | |
| Detected | 54 | 9b | 63 | PPA | 98.2% (54/55) | 90.4% to 99.7% |
| Not
Detected | 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 B
Result | Simplexa™ Flu A/B & RSV
Direct (Gen 2.0) Flu B
Result | | Grand
Total | | %Agreement | 95% CI |
|----------------------------------------------------------------------|-------------------------------------------------------------|-----------------|----------------|-----|---------------------|-----------------|
| | Detected | Not
Detected | | | | |
| Detected | 56 | 0 | 63 | PPA | 100.0% (56/56) | 93.6% to 100.0% |
| Not
Detected | 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

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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 & RSV
Direct (Gen 1.0) RSV Result | | Grand
Total | | %Agreement | 95% CI |
|--------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------|--------------|----------------|-----|-----------------|----------------|
| | Detected | Not Detected | | | | |
| Detected | 45 | 9d | 54 | PPA | 97.8% (45/46) | 88.7% to 99.6% |
| Not
Detected | 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 FDA
cleared NAT. | | | | | | |
| e1/1 discrepant (K120413 – Positive and K142365 – Negative) sample was negative for RSV on another FDA
cleared 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 & RSV
Direct (Gen 2.0) RSV Result | | Grand
Total | | %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.

OrganismConcentration TestedResult
Influenza A/Taiwan/42/06 H1N1100 TCID50/mLFlu A Detected
Influenza A/Anhui/1/2013 (H7N9)25,000 EID50/mLFlu A Detected
Influenza A/Brisbane/10/07 H3100 TCID50/mLFlu A Detected
Influenza A/Brisbane/59/07 H1100 TCID50/mLFlu A Detected
OrganismConcentration TestedResult
Influenza A/California/12/2012 (H1N1) pdm09100 TCID50/mLFlu A Detected
Influenza A/California/7/2009 NYMC x-179-A100 IU/mLFlu A Detected
Influenza A/Indiana/08/2011 (H3N2)v100 TCID50/mLFlu A Detected
Influenza A/Minnesota/11/2010 (H3N2)v100 CEID50/mLFlu A Detected
Influenza A/New Caledonia/20/99 H1N1100 TCID50/mLFlu A Detected
Influenza A/Ohio/02/2012 (H3N2)200 CEID50/mLFlu A Detected
Influenza A/Port Chalmers/1/73 H3N2100 TCID50/mLFlu A Detected
Influenza A/PR/8/34 H1N1100 TCID50/mLFlu A Detected
Influenza A/Solomon Island/03/06100 TCID50/mLFlu A Detected
Influenza A/Swine NY/02/2009 H1N1100 TCID50/mLFlu A Detected
Influenza A/Swine/1976/31 H1N1100 TCID50/mLFlu A Detected
Influenza A/Swine/lowa/15/30 H1N1100 TCID50/mLFlu A Detected
Influenza A/Texas/50/2012 (H3N2)100 TCID50/mLFlu A Detected
Influenza A/Wisconsin/67/05 H3100 TCID50/mLFlu A Detected
Influenza A/WS/33 H1N1100 TCID50/mLFlu A Detected
Influenza B/Allen/45100 TCID50/mLFlu B Detected
Influenza B/Brisbane/60/2008100 CEID50/mLFlu B Detected
Influenza B/Florida/02/2006100 TCID50/mLFlu B Detected
Influenza B/Florida/04/2006100 TCID50/mLFlu B Detected
Influenza B/Florida/07/04100 TCID50/mLFlu B Detected
Influenza B/Hong Kong/5/72100 TCID50/mLFlu B Detected
Influenza B/Lee/40100 TCID50/mLFlu B Detected
Influenza B/Maryland/1/59100 TCID50/mLFlu B Detected
Influenza B/Panama/45/90100 TCID50/mLFlu B Detected
Influenza B/Taiwan/2/62100 TCID50/mLFlu B Detected
Influenza B/Wisconsin/01/2010100 CEID50/mLFlu B Detected
RSV-A Long100 TCID50/mLRSV Detected
RSV-B 9320100 TCID50/mLRSV Detected
OrganismConcentration TestedResult
RSV-B Wash/18537/62100 TCID50/mLRSV Detected
RSV B WV/14617/85100 TCID50/mLRSV Detected

11

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K142365

510(k) Summary

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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)
OrganismConc.NFlu A (FAM)Flu B (JOE)RSV (CFR610)IC (Q670)
BaselineN/A50.0%(0/5)0.0%(0/5)0.0%(0/5)100.0%(5/5)
Adenovirus 14.17E+05
TCID50/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Adenovirus 7A5.37E+05
TCID50/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Bordetella pertussis
A6391.88E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Chlamydia
pneumoniae1.00E+06
IFU/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Cytomegalovirus
(CMV)1.04E+05
U/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Coronavirus 229E5.89E+05
TCID50/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Coronavirus OC431.95E+05
TCID50/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Corynebacterium
diphtheriae4.00E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Enterovirus Type 711.10E+05
TCID50/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Epstein-Barr Virus
(EBV)1.10E+05
copies/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Escherichia coli
0157:H71.10E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Haemophilus
influenzae1.41E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Lactobacillus
plantarum, 17-57.97E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Legionella
longbeachae8.63E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Measles1.95E+05
TCID50/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Metapneumovirus 91.58E+05
TCID50/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Moraxella catarrhalis,
NE11.49E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)

13

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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)
OrganismTested
Conc.Flu A (FAM)Flu B (JOE)RSV (CFR610)IC (Q670)
Mumps8.51E+05
TCID50/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Mycobacterium
tuberculosis (genomic
DNA)6.54E+06
c/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Mycoplasma
pneumoniae, M1293.16E+06
ccu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Neisseria elongata2.05E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Neisseria
meningitides7.07E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Parainfluenza 11.15E+05
TCID50/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Parainfluenza 23.80E+05
IU/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Parainfluenza 31.95E+05
TCID50/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Pseudomonas
aeruginosa3.93E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Rhinovirus 1A1.26E+05
TCID50/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Staphylococcus
aureus, COL1.42E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Staphylococcus
epidermidis9.23E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Streptococcus
pneumoniae9.20E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Streptococcus
pyogenes, M11.36E+06
cfu/mL30.0%(0/3)0.0%(0/3)0.0%(0/3)100.0%(3/3)
Streptococcus
salivarius2.12E+06
cfu/mL30.0%(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.

SubstanceConcentration TestedQualitative Result for each Channel
Flu A (FAM)Flu B (Joe)RSV (CFR610)RNA IC (Q670)
BaselineNone100.0%(15/15)100.0%(15/15)100.0%(15/15)100.0%(15/15)
Afrin Nasal Spray15% (v/v)100.0%(3/3)100.0%(3/3)100.0%(3/3)100.0%(3/3)
Oseltamivir phosphate1 $\mu$ M100.0%(3/3)100.0%(3/3)100.0%(3/3)100.0%(3/3)
Blood2% (v/v)100.0%(3/3)100.0%(3/3)100.0%(3/3)100.0%(3/3)
Zicam Nasal Gel5% (v/v)100.0%(3/3)100.0%(3/3)100.0%(3/3)100.0%(3/3)
Tobramycin4 $\mu$ g/mL100.0%(3/3)100.0%(3/3)100.0%(3/3)100.0%(3/3)
Mupirocin6.6 mg/mL100.0%(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 | Concentration
Tested | Qualitative Result for each Channel | | | |
|------------------------------------------|-------------------------|-------------------------------------|-------------|--------------|---------------|
| | | Flu A (FAM) | Flu B (Joe) | RSV (CFR610) | RNA IC (Q670) |
| Purified Mucin
Protein | 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) |
| Nasal
Corticosteroid

  • Fluticasone | 5% (v/v) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) |
    | Relenza
    Antiviral 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) |
    | Positive
    Control | 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 |

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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)Sample
Panel
MemberNMean
CtInter-DayInter-RunInter-LotIntra-
Run/LotTotal
SD%CVSD%CVSD%CVSD%CVSD%CV
Flu A
(FAM)Flu A Low
Positive3637000.30.9000.71.90.82.1
Flu A
Moderate
Positive3635.200000.410.41.10.51.5
Positive
Control
(PC)3633.30.10.30.20.50.20.50.20.50.31
Flu B
(JOE)Flu B Low
Positive3635.300000.30.80.71.90.72.1
Flu B
Moderate
Positive4133.5000.10.40.30.90.310.51.4
Positive
Control
(PC)3632.700000.41.10.20.70.41.3
RSV
(CFR610)RSV Low
Positive3636.50000000.82.30.82.3
RSV
Moderate
Positive3634.6000.20.6000.51.40.51.5
Positive
Control
(PC)3632.40.10.40.10.200.10.20.60.20.7
RNA IC
(Q670)Flu A Low
Positive362900000.41.50.41.20.61.9
Flu A
Moderate
Positive3629.100000.41.40.41.20.61.9
Flu B Low
Positive3628.20.20.5000.20.80.310.41.4
Flu B
Moderate
Positive4128.200.100.10.20.70.20.70.31
RSV Low
Positive3628.900000.20.80.31.10.41.3
RSV
Moderate
Positive3628.900000.30.90.30.90.41.2
Negative3629.2000.10.2000.310.31
Positive
Control3628.900.1000.20.70.10.40.20.8

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) | Sample
Panel
Member | 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 | | | | | |

| Qualitative
Results

(By Sample)AllFlu A (FAM)Flu B (JOE)RSV (CFR610)
Detected
NNot Detected
NDetected
NNot Detected
NDetected
NNot Detected
N
%%%%%%
Flu A Low
Positive36363636
100.0 %100.0 %100.0 %
Flu A
Medium
Positive36363636
100.0 %100.0 %100.0 %
Flu B Low
Positive36363636
100.0 %100.0 %100.0 %
Flu B
Medium
Positive41414141
100.0 %100.0 %100.0 %
RSV: Low
Positive36363636
100.0 %100.0 %100.0 %
RSV:
Medium
Positive36363636
100.0 %100.0 %100.0 %
Negative
Template
Control
(NTC)36363636
100.0 %100.0 %100.0 %
Positive
Control
(PC)36363636
100.0 %100.0 %100.0 %

ADDITIONAL PERFORMANCE STUDIES

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