Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

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

Therapeutic

No.
The device is an in vitro diagnostic (IVD) assay designed for the qualitative detection and differentiation of HSV-1 and HSV-2 DNA, which is used for diagnosis, not treatment.

Diagnostic

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states, "This test is an aid in the differential diagnosis of HSV-1 and HSV-2 genital infections," which directly indicates its use for diagnostic purposes.

SaMD (Software as a Medical Device)

No

The device description explicitly states that the system consists of the Simplexa™ HSV 1 & 2 Direct assay, the 3M Integrated Cycler (with 3M Integrated Cycler Studio Software), the Direct Amplification Disc and associated accessories. This includes hardware components (3M Integrated Cycler, Direct Amplification Disc, accessories) in addition to software.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The intended use explicitly states it is for the "qualitative detection and differentiation of herpes simples virus (HSV-1 and HSV-2) DNA present in genital lesion swabs samples from patients with signs and symptoms of HSV-1 or HSV-2 infection of the genitalia." This is a diagnostic purpose performed on a biological sample (genital lesion swabs).
Device Description: The description details a "real-time PCR" system that analyzes "unprocessed genital swab specimens." PCR is a common technique used in in vitro diagnostics to detect genetic material from pathogens.
Performance Studies: The document includes detailed performance studies (Prospective and Retrospective Clinical Agreement, Reproducibility, Analytical Sensitivity, Analytical Reactivity, Cross-Reactivity, Interference) which are standard for demonstrating the performance of an IVD.
Key Metrics: The document provides key metrics like Sensitivity, Specificity, PPV, and NPV, which are used to evaluate the clinical performance of diagnostic tests.
Predicate Device: The mention of a "Predicate Device(s)" with a K number (K142738; artus® HSV-1/2 QS-RGQ MDx Kit) indicates that this device is being compared to a previously cleared IVD, a common process for regulatory submission of new IVDs.

All of these factors strongly indicate that the Simplexa™ HSV 1 & 2 Direct assay is an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

Simplexa™ HSV 1 & 2 Direct

The Focus Diagnostics Simplexa™ HSV 1 & 2 Direct assay is intended for use on the 3M Integrated Cycler instrument for the qualitative detection and differentiation of herpes simples virus (HSV-2) DNA present in genital lesion swabs samples from patients with signs and symptoms of HSV-2 infection of the genitalia. This test is an aid in the differential diagnosis of HSV-1 and HSV-2 genital infections.

The assay is not intended for use as a screening test for the presence of HSV-2 in blood or blood products. The assay is for professional use only.

Simplexa™ HSV 1 & 2 Positive Control Pack

The Simplexa™ HSV 1 & 2 Positive Control Pack is intended to be used as a control with the Simplexa™ HSV 1 & 2 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)

OQO

Device Description

The Simplexa™ HSV 1 & 2 Direct assay system is a real-time PCR that enables the direct amplification, detection and differentiation of HSV-1 and/or HSV-2 DNA from unprocessed genital swab specimens without nucleic acid extraction. The system consists of the Simplexa™ HSV 1 & 2 Direct assay, the 3M Integrated Cycler (with 3M Integrated Cycler Studio Software), the Direct Amplification Disc and associated accessories.

In the Simplexa™ HSV 1 & 2 Direct assay, bi-functional fluorescent probe-primers are used together with corresponding reverse primers to amplify HSV-1, HSV-2 and internal control targets. Well conserved regions of the HSV-1 and HSV-2 DNA polymerase genes are targeted to identify HSV-1 and HSV-2 DNA respectively in the specimen. An internal control is used to detect PCR failure and/or inhibition.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

genital lesion swabs samples

Indicated Patient Age Range

17 Years of age or older

Intended User / Care Setting

professional use only.

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

Reproducibility: Evaluated at three investigative sites. Each laboratory tested one positive control and a panel of five contrived sample pools (low and medium positive for each analyte, and a high negative). Assays were performed in triplicate on five different days. Each site had two operators, each assaying the entire sample panel and positive control once per day.

Analytical Sensitivity/Limit of Detection (LoD): Determined using quantified stocks of HSV-1 and HSV-2 serially diluted into negative genital swab matrix. LoD was the lowest concentration detected positive > 95% of the time.

Analytical Reactivity: Evaluated using different HSV-1 and HSV-2 strains not used for LoD determination. Quantified viral material spiked into negative genital swab matrix (male and female genital swabs) at a single dilution and assayed in triplicate.

Cross-Reactivity (Analytical Specificity): Evaluated by testing against 36 potential cross-reactants spiked into negative genital swab matrix (male and female genital swabs) and assayed in triplicate.

Interference: Evaluated with 26 potentially interfering substances at specified concentrations, tested in a low positive HSV-1 and HSV-2 sample (4 times LoD) in negative genital swab matrix (male and female genital swabs) and assayed in triplicate.

Clinical Agreement - Prospective Study:
Sample Size: 696 genital swab samples.
Data Source: Prospectively collected from patients with signs and symptoms of genital herpes simplex virus (HSV) infection from 6 geographically diverse locations.
Annotation Protocol: Compared to a composite comparator algorithm consisting of culture, bi-directional sequencing, and an FDA-cleared NAAT. A positive result for HSV-1 and/or HSV-2 was determined by a positive test result in either culture or bi-directional sequencing. If both were positive but disagreed on differentiation, results of the FDA-cleared NAAT were used; a "2 out of 3" rule determined the final type. Samples collected on the Simplexa™ HSV-1 and HSV-2 Direct and sent to a central lab for culture testing (HSV-2 first, then HSV-1 if negative). Retained samples were sent to Focus Diagnostics for bi-directional sequencing.

Clinical Agreement - Retrospective Study:
Sample Size: 28 genital swab samples (14 positive HSV-1 and 14 positive HSV-2).
Data Source: Retrospectively collected from male patients with signs and symptoms of genital herpes simplex virus (HSV) infection; preselected positive and negative samples.
Annotation Protocol: Tested at Focus Diagnostics using the Simplexa™ HSV 1 & 2 Direct and a validated bi-directional sequencing assay.

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

Reproducibility - Genital Swab Sample Type
Study type: Reproducibility study.
Sample size: Multiple samples tested in triplicate on five different days across three sites. (Total 539 results used for total agreement calculation)
Key Results:
HSV-1 Result:
HSV-1 Low Positive: 100.0% (90/90) agreement with expected results.
HSV-1 Medium Positive: 100.0% (90/90) agreement with expected results.
HSV-2 Low Positive: 98.9% (89/90) agreement with expected results (Negative for HSV-1).
HSV-2 Medium Positive: 98.9% (89/90) agreement with expected results (Negative for HSV-1).
High Negative: 93.3% (84/90) agreement with expected results (Negative for HSV-1).
Positive Control: 100.0% (89/89) agreement with expected results.
Total Agreement for HSV-1 Result: 98.5% (531/539).

HSV-2 Result:
HSV-1 Low Positive: 98.9% (89/90) agreement with expected results (Negative for HSV-2).
HSV-1 Medium Positive: 100.0% (90/90) agreement with expected results (Negative for HSV-2).
HSV-2 Low Positive: 94.4% (85/90) agreement with expected results.
HSV-2 Medium Positive: 100.0% (90/90) agreement with expected results.
High Negative: 94.4% (85/90) agreement with expected results (Negative for HSV-2).
Positive Control: 100.0% (89/89) agreement with expected results.
Total Agreement for HSV-2 Result: 98.0% (528/539).

DNA IC Result (Internal Control):
All sample types (HSV-1 Low Positive, HSV-1 Medium Positive, HSV-2 Low Positive, HSV-2 Medium Positive, High Negative, Positive Control) showed 100.0% agreement with expected results for the internal control.
Total Agreement for DNA IC Result: 100.0% (539/539).

Analytical Sensitivity/Limit of Detection (LoD) - Genital Swab Sample Type
Study type: Analytical sensitivity determination.
Sample size: 32 replicates per virus strain.
Key Results:
HSV-1 McIntyre: LoD Concentration 4 TCID50/mL, 32/32 detected.
HSV-1 HF: LoD Concentration 160 TCID50/mL, 32/32 detected.
HSV-2 G: LoD Concentration 2 TCID50/mL, 32/32 detected.
HSV-2 MS: LoD Concentration 10 TCID50/mL, 31/32 detected.

Analytical Reactivity - Genital Swab Sample Type
Study type: Analytical reactivity assessment.
Sample size: 3 replicates per strain/isolate.
Key Results: The Simplexa™ HSV 1 & 2 Direct assay was able to detect other strains of HSV-1 and HSV-2 viruses not used in LoD determination.
HSV-1 KOS: 3/3 detected for HSV-1, 0/3 detected for HSV-2.
HSV-1 F: 3/3 detected for HSV-1, 0/3 detected for HSV-2.
HSV-2 Isolate 1: 0/3 detected for HSV-1, 3/3 detected for HSV-2.
HSV-2 Isolate 2: 0/3 detected for HSV-1, 3/3 detected for HSV-2.
HSV-2 Isolate 3: 0/3 detected for HSV-1, 3/3 detected for HSV-2.

Cross-Reactivity (Analytical Specificity) - Genital Swab Sample Type
Study type: Analytical specificity assessment.
Sample size: 3 replicates for each of 36 potential cross-reactants.
Key Results: No cross-reactivity was observed for any of the tested microorganisms/viruses, except for Enterovirus 71 (1/8 detected for HSV-2).

Interference - Genital Swab Sample Type
Study type: Interference study.
Sample size: 3 replicates per interferent (some noted exceptions).
Key Results: No interference was observed for most tested substances. One out of three (1/3) initial replicates and two out of five (2/5) confirmation replicates were negative for HSV-2 when Acyclovir Cream was tested as a potential interferent.

CLINICAL AGREEMENT - Genital Swab Sample Type

Prospective Study
Study type: Clinical Agreement Study
Sample size: 696 samples
Key Results:
Simplexa™ HSV 1 & 2 Direct Compared to Composite Comparator Result (HSV-1)
Sensitivity: 97.4%(111/114)
Specificity: 98.2%(560/570)
PPV: 91.7%(111/121)
NPV: 99.5%(560/563)

Simplexa™ HSV 1 & 2 Direct Compared to Composite Comparator Result (HSV-2)
Sensitivity: 97.2%(175/180)
Specificity: 97.8%(497/508)
PPV: 94.1%(175/186)
NPV: 99.0%(497/502)

Retrospective Study
Study type: Clinical Agreement Study
Sample size: 28 samples
Key Results:
HSV-1 Validated Bi-Directional Sequencing Assay Results
PPA: 100.0%(14/14)
NPA: 92.9%(13/14)
PPV: 93.3%(14/15)
NPV: 100.0%(13/13)

HSV-2 Validated Bi-Directional Sequencing Assay Results
PPA: 100.0%(14/14)
NPA: 100.0%(14/14)
PPV: 100.0%(14/14)
NPV: 100.0%(14/14)

FRESH VS FROZEN STUDY
Study type: Storage conditions validation.
Key Results: Samples should be transported on ice and stored at 2 to 8°C for up to 7 days post collection. If there is a greater than 7 day delay before processing, store the sample at -70° C.

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

Prospective Study:
HSV-1:
Sensitivity: 97.4% (111/114), 95% CI: 92.5% to 99.1%
Specificity: 98.2% (560/570), 95% CI: 96.8% to 99.0%
PPV: 91.7% (111/121), 95% CI: 85.5% to 95.4%
NPV: 99.5% (560/563), 95% CI: 98.4% to 99.8%

HSV-2:
Sensitivity: 97.2% (175/180), 95% CI: 93.7% to 98.8%
Specificity: 97.8% (497/508), 95% CI: 96.2% to 98.8%
PPV: 94.1% (175/186), 95% CI: 89.7% to 96.7%
NPV: 99.0% (497/502), 95% CI: 97.7% to 99.6%

Retrospective Study:
HSV-1:
PPA: 100.0% (14/14), 95% CI: 78.5% to 100.0%
NPA: 92.9% (13/14), 95% CI: 68.5% to 98.7%
PPV: 93.3% (14/15), 95% CI: 70.2% to 98.8%
NPV: 100.0% (13/13), 95% CI: 77.2% to 100.0%

HSV-2:
PPA: 100.0% (14/14), 95% CI: 78.5% to 100.0%
NPA: 100.0% (14/14), 95% CI: 78.5% to 100.0%
PPV: 100.0% (14/14), 95% CI: 78.5% to 100.0%
NPV: 100.0% (14/14), 95% CI: 78.5% to 100.0%

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.

K142738

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

Not Found

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

Not Found

Regulation Number and Section

§ 866.3305 Herpes simplex virus serological assays.

(a)
Identification. Herpes simplex virus serological assays are devices that consist of antigens and antisera used in various serological tests to identify antibodies to herpes simplex virus in serum. Additionally, some of the assays consist of herpes simplex virus antisera conjugated with a fluorescent dye (immunofluorescent assays) used to identify herpes simplex virus directly from clinical specimens or tissue culture isolates derived from clinical specimens. The identification aids in the diagnosis of diseases caused by herpes simplex viruses and provides epidemiological information on these diseases. Herpes simplex viral infections range from common and mild lesions of the skin and mucous membranes to a severe form of encephalitis (inflammation of the brain). Neonatal herpes virus infections range from a mild infection to a severe generalized disease with a fatal outcome.(b)
Classification. Class II (special controls). The device is classified as class II (special controls). The special control for the device is FDA's revised guidance document entitled “Class II Special Controls Guidance Document: Herpes Simplex Virus Types 1 and 2 Serological Assays.” For availability of the guidance revised document, see § 866.1(e).

Summary

0

Image /page/0/Picture/1 description: The image is a black and white logo for the U.S. Department of Health and Human Services. The logo features a stylized image of an eagle with three curved lines representing its wings. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the eagle image. The text is in all caps and is evenly spaced around the circle.

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

August 28, 2015

FOCUS DIAGNOSTICS, INC. SHARON YOUNG SENIOR REGULATORY AFFAIRS SPECIALIST 11331 VALLEY VIEW STREET CYPRESS, CA 90630

Re: K150962

Trade/Device Name: Simplexa HSV 1 & 2 Direct, Simplexa HSV1 & 2 Positive Control Pack

Regulation Number: 21 CFR 866.3305 Regulation Name: Herpes simplex virus serological assays Regulatory Class: II Product Code: OQO Dated: August 25, 2015 Received: August 26, 2015

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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

1

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), 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" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of 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,

Stephen J. Lovell -S for

Uwe Scherf, 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) K150962

Device Name

Simplexa™ HSV 1 & 2 Direct and Simplexa™ HSV 1 & 2 Positive Control Pack

Indications for Use (Describe)

Simplexa™ HSV 1 & 2 Direct

The Focus Diagnostics Simplexa™ HSV 1 & 2 Direct assay is intended for use on the 3M Integrated Cycler instrument for the qualitative detection and differentiation of herpes simples virus (HSV-2) DNA present in genital lesion swabs samples from patients with signs and symptoms of HSV-2 infection of the genitalia. This test is an aid in the differential diagnosis of HSV-1 and HSV-2 genital infections.

The assay is not intended for use as a screening test for the presence of HSV-2 in blood or blood products. The assay is for professional use only.

Simplexa™ HSV 1 & 2 Positive Control Pack

The Simplexa™ HSV 1 & 2 Positive Control Pack is intended to be used as a control with the Simplexa™ HSV 1 & 2 Direct kit. This control is not intended for use with other assays or systems.

X Prescription Use (Part 21 CFR 801 Subpart D)

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

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3

Image /page/3/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, sans-serif font. A curved, black shape is to the left of the word, resembling a stylized check mark or swoosh. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin black line.

510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 1 of 12

| 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 | August 20, 2015 |
| Proprietary Name | Simplexa™ HSV 1 & 2 Direct
Simplexa™ HSV 1 & 2 Positive Control Pack |
| Generic Name | Herpes Simplex Virus serological assays |
| Classification | OQO |
| Regulation | |
| Product Code | |
| Predicate Device | OQO - artus® HSV-1/2 QS-RGQ MDx Kit (K142738) |

Intended Use

Simplexa™ HSV 1 & 2 Direct

The Focus Diagnostics Simplexa™ HSV 1 & 2 Direct assay is intended for use on the 3M Integrated Cycler instrument for the qualitative detection and differentiation of herpes simples virus (HSV-1 and HSV-2) DNA present in genital lesion swabs samples from patients with signs and symptoms of HSV-1 or HSV-2 infection of the genitalia. This test is an aid in the differential diagnosis of HSV-1 and HSV-2 genital infections.

The assay is not intended for use as a screening test for the presence of HSV-1 and HSV-2 in blood or blood products. The assay is for professional use only.

Simplexa™ HSV 1 & 2 Positive Control Pack

The Simplexa™ HSV 1 & 2 Positive Control Pack is intended to be used as a control with the Simplexa™ HSV 1 & 2 Direct kit. This control is not intended for use with other assays or systems.

Device Description

The Simplexa™ HSV 1 & 2 Direct assay system is a real-time PCR that enables the direct amplification, detection and differentiation of HSV-1 and/or HSV-2 DNA from unprocessed genital swab specimens without nucleic acid extraction. The system consists of the Simplexa™ HSV 1 & 2 Direct assay, the 3M Integrated Cycler (with 3M Integrated Cycler Studio Software), the Direct Amplification Disc and associated accessories.

In the Simplexa™ HSV 1 & 2 Direct assay, bi-functional fluorescent probe-primers are used together with corresponding reverse primers to amplify HSV-1, HSV-2 and internal control targets. Well conserved regions of the HSV-1 and HSV-2 DNA polymerase genes are targeted to identify HSV-1 and HSV-2 DNA respectively in the specimen. An internal control is used to detect PCR failure and/or inhibition.

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Image /page/4/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of a stylized swoosh shape on the left, followed by the word "FOCUS" in a bold, sans-serif font. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin line. The entire logo is in black and white.

510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 2 of 12

Predicate Device Information

Similarities

| Name | Simplexa™ HSV
1 & 2 Direct | artus® HSV-1/2 QS-RGQ MDx Kit
K142738 |
|-------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | Simplexa™ HSV 1 & 2 Direct
The Focus Diagnostics Simplexa™ HSV 1 &
2 Direct assay is intended for use on the 3M
Integrated Cycler instrument for the
qualitative detection and differentiation of
herpes simplex virus (HSV-1 and HSV-2)
DNA present in genital lesion swabs samples
from patients with signs and symptoms of
HSV-1 or HSV-2 infection of the genitalia.
This test is an aid in the differential diagnosis
of HSV-1 and HSV-2 genital infections.
The assay is not intended for use as a
screening test for the presence of HSV-1 and
HSV-2 in blood or blood products. The assay
is for professional use only.

Simplexa™ HSV 1 & 2 Positive Control Pack
The Simplexa™ HSV 1 & 2 Positive Control
Pack is intended to be used as a control with
the Simplexa™ HSV 1 & 2 Direct kit.

This control is not intended for use with other
assays or systems. | The artus HSV-1/2 QS-RGQ MDx Kit is an in vitro
real-time PCR DNA amplification assay performed
on the QIAsymphony RGQ MDx system for the
direct qualitative detection and differentiation of
herpes simplex virus (HSV-1 and HSV-2) DNA in
genital or oral vesicular lesions from male and
female patients suspected of HSV infection.
The assay is intended for use as an aid in diagnosis
of HSV infection in symptomatic patients.
Warning: The artus HSV-1/2 QS-RGQ MDx Kit is
not FDA-cleared for use with cerebrospinal fluid
(CSF) or for prenatal screening. |
| Assay
Targets | HSV-1 and HSV-2 DNA | HSV-1 and HSV-2 DNA |
| Detection
Techniques | Multiplex assay using different reporter dyes
for each target. | Multiplex assay using different reporter dyes for
each target. |

Differences

| Name | Simplexa™ HSV
1 & 2 Direct | artus® HSV-1/2 QS-RGQ MDx Kit
K142738 |
|-------------------|------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Sample Types | Genital herpetic lesion swab samples | Male and female genital or oral herpetic lesions. |
| Assay Methodology | PCR-based system for detecting the
presence / absence of viral DNA in clinical
specimens | Amplification and detection are carried out using the
artus HSV-1/2 QS-RGQ MDx Kit with the Rotor-
Gene Q MDx (RGQ MDx) and Rotor-Gene Assay
Manager software. |

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 a curved, swooping line above it. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin line.

510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 3 of 12

REPRODUCIBILITY - Genital Swab Sample Type

Reproducibility for the Simplexa™ HSV 1 & 2 Direct assay was evaluated. Three investigative sites assessed the device's inter-site, inter-day and inter/intra-assay reproducibility. Each of the laboratories tested the positive control and a panel of five contrived sample pools including a low (approximately 1-2 times LoD) and medium positive (approximately 2-4 times LoD) for each analyte and a high negative. The high negative sample contained a small amount of HSV-1 and it was designed to be neqative approximately 95% of the time. The assays were performed in triplicate on five different days. Each site had two operators; each operator assayed the entire sample panel and positive control once per day, for a total of two sets of data per day. Combined results for all sites are presented in the tables below.

| | Sample | Site - 1 | | | Site - 2 | | | Site - 3 | | | Total %
Agreement
with
Expected
Results | 95% CI |
|----------------------------------------------------------------------------------------------------------------------|-----------------------------|-----------------------------------------------|------------|--------------|-----------------------------------------------|------------|--------------|-----------------------------------------------|------------|--------------|-----------------------------------------------------|-------------------|
| | | %
Agreement
with
Expected
Results | Avg.
Ct | Total
%CV | %
Agreement
with
Expected
Results | Avg.
Ct | Total
%CV | %
Agreement
with
Expected
Results | Avg.
Ct | Total
%CV | | |
| | HSV-1 Low
Positive | 100.0%
(30/30) | 36.0 | 2.2 | 100.0%
(30/30) | 36.1 | 2.6 | 100.0%
(30/30) | 36.3 | 2.7 | 100.0%
(90/90) | 95.9 to
100.0% |
| HSV-1
Result | HSV-1
Medium
Positive | 100.0%
(30/30) | 34.4 | 1.7 | 100.0%
(30/30) | 34.8 | 1.2 | 100.0%
(30/30) | 34.6 | 1.9 | 100.0%
(90/90) | 95.9 to
100.0% |
| | HSV-2 Low
Positive | 100.0%
(30/30)a | NA | NA | 100.0%
(30/30)a | NA | NA | 96.7%
(29/30)a | NA | NA | 98.9%
(89/90)a | 94.0 to
99.8% |
| | HSV-2
Medium
Positive | 100.0%
(30/30)a | NA | NA | 96.7%
(29/30)a | NA | NA | 100.0%
(30/30)a | NA | NA | 98.9%
(89/90)a | 94.0 to
99.8% |
| | High
Negative | 96.7%
(29/30)a | 38.8 | 0.0 | 93.3%
(28/30)a | 38.7 | 0.5 | 90.0%
(27/30)a | 38.0 | 4.1 | 93.3%
(84/90)a | 86.2 to
96.9% |
| | Positive
Control | 100.0%
(30/30) | 29.9 | 0.8 | 100.0%
(30/30) | 30.4 | 1.3 | 100.0%
(29/29) | 29.9 | 2.8 | 100.0%
(89/89) | 95.9 to
100.0% |
| | Total
Agreement | 99.4% (179/180) | | | 98.3% (177/180) | | | 97.8% (175/179) | | | 98.5%
(531/539) | 97.1 to
99.2% |
| a) Expected Results of HSV-2 Low Positive, HSV-2 Medium Positive and High Negative samples are "Negative" for HSV-1. | | | | | | | | | | | | |

6

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510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 4 of 12

| | Sample | Site - 1 | | | Site - 2 | | | Site - 3 | | | Total %
Agreement
with
Expected
Results | 95% CI |
|-----------------|----------------------------------------------------------------------------------------------------------------------|-----------------------------------------------|------------|-----------------|-----------------------------------------------|------------|-----------------|-----------------------------------------------|------------|--------------------|-----------------------------------------------------|-------------------|
| | | %
Agreement
with
Expected
Results | Avg.
Ct | Total
%CV | %
Agreement
with
Expected
Results | Avg.
Ct | Total
%CV | %
Agreement
with
Expected
Results | Avg.
Ct | Total
%CV | | |
| HSV-2
Result | HSV-1
Low
Positive | 100.0%
(30/30)b | NA | NA | 100.0%
(30/30)b | NA | NA | 96.7%
(29/30)b | 41.1 | 0.0 | 98.9%
(89/90)b | 94.0 to
99.8% |
| | HSV-1
Medium
Positive | 100.0%
(30/30)b | NA | NA | 100.0%
(30/30)b | NA | NA | 100.0%
(30/30)b | NA | NA | 100.0%
(90/90)b | 95.9 to
100.0% |
| | HSV-2
Low
Positive | 100.0%
(30/30) | 37.4 | 2.9 | 90.0%
(27/30) | 37.5 | 3.5 | 93.3%
(28/30) | 37.1 | 2.8 | 94.4%
(85/90) | 87.6 to
97.6% |
| | HSV-2
Medium
Positive | 100.0%
(30/30) | 35.5 | 1.9 | 100.0%
(30/30) | 35.6 | 2.0 | 100.0%
(30/30) | 35.3 | 1.6 | 100.0%
(90/90) | 95.9 to
100.0% |
| | High
Negative | 96.7%
(29/30)b | 39.5 | 0.0 | 86.7%
(26/30)b | 38.6 | 2.9 | 100.0%
(30/30)b | NA | NA | 94.4%
(85/90)b | 87.6 to
97.6% |
| | Positive
Control | 100.0%
(30/30) | 30.2 | 1.3 | 100.0%
(30/30) | 30.1 | 0.6 | 100.0%
(29/29) | 29.9 | 1.2 | 100.0%
(89/89) | 95.9 to
100.0% |
| | Total
Agreement | 99.4% (179/180) | | 96.1% (173/180) | | | 98.9% (176/179) | | | 98.0%
(528/539) | 96.4 to
98.9% | |
| | b) Expected Results of HSV-1 Low Positive, HSV-1 Medium Positive and High Negative samples are "Negative" for HSV-2. | | | | | | | | | | | |

7

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510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 5 of 12

	Sample	Site - 1
	Sample	% Agreement with Expected Results	Avg.
Ct	Total
%CV	% Agreement with Expected Results	Avg.
Ct	Total
%CV	% Agreement with Expected Results	Avg.
Ct	Total
%CV	Total % Agreement with Expected Results	95% CI
DNA
IC
Result	HSV-1 Low
Positive	100.0%
(30/30)	29.6	0.7	100.0%
(30/30)	29.8	1.2	100.0%
(30/30)	29.7	1.0	100.0%
(90/90)	95.9 to
100.0%
	HSV-1
Medium
Positive	100.0%
(30/30)	29.6	0.8	100.0%
(30/30)	29.8	1.4	100.0%
(30/30)	29.7	0.9	100.0%
(90/90)	95.9 to
100.0%
	HSV-2 Low
Positive	100.0%
(30/30)	29.6	0.8	100.0%
(30/30)	29.8	1.2	100.0%
(30/30)	29.7	1.0	100.0%
(90/90)	95.9 to
100.0%
	HSV-2
Medium
Positive	100.0%
(30/30)	29.5	0.6	100.0%
(30/30)	29.7	1.4	100.0%
(30/30)	29.8	1.4	100.0%
(90/90)	95.9 to
100.0%
	High
Negative	100.0%
(30/30)	29.6	0.6	100.0%
(30/30)	29.8	1.2	100.0%
(30/30)	29.7	1.0	100.0%
(90/90)	95.9 to
100.0%
	Positive
Control	100.0%
(30/30)	29.5	0.5	100.0%
(30/30)	29.7	1.4	100.0%
(29/29)	29.7	0.9	100.0%
(89/89)	95.9 to
100.0%
	Total
Agreement	100.0% (180/180)		100.0% (180/180)	100.0% (179/179)	100.0%
(539/539)	96.4 to
98.9%

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION - Genital Swab Sample Type

The Limit of Detection (LoD) was determined for the Simplexa™ HSV 1 & 2 Direct assay using quantified
stocks of HSV-1 and HSV-2 serially diluted into negative genital swab ma genital swabs. LoD was determined to be the lowest concentration that could be detected positive > 95% of the time.

| Virus Strain | LoD Concentration
(TCID50/mL) | Qualitative
Results
(#Detected/#Total) | Mean Ct ± SD
(from Detected
Replicates only) |
|----------------|----------------------------------|----------------------------------------------|----------------------------------------------------|
| HSV-1 McIntyre | 4 | 32/32 | 36.4 ± 1.16 |
| HSV-1 HF | 160 | 32/32 | 35.2 ± 1.03 |
| HSV-2 G | 2 | 32/32 | 37.5 ± 1.08 |
| HSV-2 MS | 10 | 31/32 | 37.9 ± 1.15 |

8

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510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 6 of 12

ANALYTICAL REACTIVITY/ CROSS REACTIVITY

Analytical Reactivity -Genital Swab Sample Type

The analytical reactivity of the Simplexa™ HSV 1 & 2 Direct assay was evaluated using different strains of HSV-1 and HSV-2 that were not used in the determination of the limit of detection (LoD) for the assay. Quantified viral material was spiked into negative genital swab matrix containing male and female genital swabs using a single dilution and assayed in triplicate. The Simplexa™ HSV 1 & 2 Direct assay was able to detect other strains of HSV-1 and HSV-2 viruses.

| HSV
Strain/Isolate | Spiked Concentration
[TCID50/mL] | Qualitative Result
(#Detected/#Total) | |
|-----------------------|-------------------------------------|------------------------------------------|-------|
| | | HSV-1 | HSV-2 |
| HSV-1 KOS | 16 | 3/3 | 0/3 |
| HSV-1 F | 32 | 3/3 | 0/3 |
| HSV-2 Isolate 1 | 8 | 0/3 | 3/3 |
| HSV-2 Isolate 2 | 8 | 0/3 | 3/3 |
| HSV-2 Isolate 3 | 8 | 0/3 | 3/3 |

Cross-Reactivity (Analytical Specificity) - Genital Swab Sample Type

The Simplexa™ HSV 1 & 2 Direct assay's analytical specificity was evaluated by testing the ability to exclusively identify HSV-1 and HSV-2 viruses with no cross-reactivity to organisms that are closely related, or cause similar clinical symptoms or may be present on swabs of the genital region. Thirty six (36) potential cross-reactants were spiked into negative genital swab matrix containing male and female genital swabs and assayed in triplicate. No cross-reactivity was observed.

| No. | Potential Cross-Reactants (#Detected/#Total) | |-----|---------------------- | | | 1 | None (Baseline) | 2 | Bacteroides fragilis | 3 | Bacteroides ureolyticus* | 4 | Candida albicans | 5 | Chlamydia trachomatis | 6 | Clostridium sordellii | 7 | Corynebacterium genitalium | 8 | Cytomegalovirus | 9 | Enterococcus faecalis vanB | 10 | Enterovirus 71 | 11 | Epstein Barr Virus (B95-8) | 12 | Escherichia coli O157H7 | 13 | Gardnerella vaginalis | 14 | Hepatitis B | 15 | Hepatitis C | 16 | HHV-6 (Z29 Strain) | 17 | HHV-7 SB | 18 | HIV-1 IIIB | Tested Concentration | Qualitative Result
|
--------|----------------------|------------------------------------------|----------------|
| | HSV-1 | HSV-2 |
| Not Applicable | 0/15 | 0/15 |
| 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| Not Applicable | Not Applicable | Not Applicable |
| 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 1.00 X 106 IFU/mL | 0/3 | 0/3 |
| 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 1.00 X 106 cfu/mL | 0/3 | 0/3 |
AD169 strain | 1.00 X 105 TCID50/mL | 0/3 | 0/3 |
| 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 1.00 X 105 TCID50/mL | 0/8 | 1/8 |
| 1.00 X 105 copies/mL | 0/3 | 0/3 |
| 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 1.00 X 105 IU/mL | 0/3 | 0/3 |
| 1.00 X 105 IU/mL | 0/3 | 0/3 |
| 1.00 X 105 TCID50/mL | 0/3 | 0/3 |
| 1.00 X 105 TCID50/mL | 0/3 | 0/3 |
| 1.00 X 105 copies/mL | 0/3 | 0/3 |

9

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510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 7 of 12

| No. | Potential Cross-Reactants | Tested Concentration | Qualitative Result
(#Detected/#Total) | |
|-----|--------------------------------------------------|----------------------------|------------------------------------------|----------------|
| | | | HSV-1 | HSV-2 |
| 19 | HIV-2 NIHZ** | Not Available | 0/3 | 0/3 |
| 20 | HPV18 Recombinant | 1.00 X 105 pfu/mL | 0/3 | 0/3 |
| 21 | Lactobacillus acidophilus | 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 22 | Mobiluncus mulieris | 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 23 | Mycoplasma genitalium** | Not Applicable | Not Applicable | Not Applicable |
| 24 | Mycoplasma hominis | 1.00 X 106 CCU/mL | 0/3 | 0/3 |
| 25 | Neisseria gonorrhoeae | 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 26 | Proteus vulgaris | 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 27 | Rubella | 1.00 X 105 TCID50/mL | 0/3 | 0/3 |
| 28 | Staphylococcus aureus (MRSA),
ATCC 700699 | 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 29 | Staphylococcus epidermidis (MRSE),
ATCC 29887 | 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 30 | Staphylococcus saprophyticus | 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 31 | Streptococcus mitis | 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 32 | Streptococcus pyogenes, M1 | 1.00 X 106 cfu/mL | 0/3 | 0/3 |
| 33 | Toxoplasma gondii | 1.00 X 106 tachyzooites/mL | 0/3 | 0/3 |
| 34 | Treponema pallidum** | Not Applicable | Not Applicable | Not Applicable |
| 35 | Trichomonas vaginalis | 1.00 X 106 trophozoites/ml | 0/3 | 0/3 |
| 36 | Ureaplasma urealyticum | 1.00 X 106 CCU/mL | 0/3 | 0/3 |
| 37 | VZV | 1.00 X 105 copies/mL | 0/3 | 0/3 |

" Microorganism was not available for testing therefore in-silico NCBI BLAST analysis was performed and found no predicted cross reactivity. ** Quantified material was not available to test; instead the vendor provided a culture fluid with a known Ct value. The site was directed to dilute the stock to a relevant Ct value; 1:50 dilution factor.

INTERFERENCE - Genital Swab Sample Type

The performance of the Simplexa™ HSV 1 & 2 Direct assay was evaluated with potentially interfering substances that may be present on swabs of the genital region at the concentrations indicated in the table below. A total of five (26) potentially interfering substances were tested in a low positive HSV-1 and HSV-2 sample (4 times LoD) in negative genital swab matrix containing male and female genital swabs and assayed in triplicate. No interference was observed.

Potential Interferent	Interferent Concentration	#Detected/#Total
		HSV-1	HSV-2	IC
Abreva cold sore treatment	7% w/v	3/3	3/3	3/3
Acyclovir	2.5 mg/mL	3/3	3/3	3/3
Acyclovir Cream*	7% w/v	8/8	5/8	8/8
Albumin	10 mg/mL	3/3	3/3	3/3
Balneol Hygienic Cleansing lotion	7% w/v	3/3	3/3	3/3
Casein	10 mg/mL	3/3	3/3	3/3
Cidofovir	2.5 mg/mL	3/3	3/3	3/3
Clotrimazole vaginal cream	7% w/v	3/3	3/3	3/3
Denavir	2.5 mg/mL	3/3	3/3	3/3

10

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510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 8 of 12

		#Detected/#Total
Potential Interferent	Interferent Concentration	HSV-1	HSV-2	IC
Douche	7% w/v	3/3	3/3	3/3
Famciclovir	2.5 mg/mL	3/3	3/3	3/3
Feces	2.5 mg/mL	3/3	3/3	3/3
Gynol II (Contraceptive jelly)	7% w/v	3/3	3/3	3/3
KY Jelly	5% v/v	3/3	3/3	3/3
Monistat 1	7% w/v	3/3	3/3	3/3
Monistat 3	7% w/v	3/3	3/3	3/3
Mucin	7% w/v	3/3	3/3	3/3
Preparation H Hemorrhoid cream	7% w/v	3/3	3/3	3/3
Releev cold sore treatment	7% w/v	3/3	3/3	3/3
Urine	10% v/v	3/3	3/3	3/3
Vagicaine Anti-Itch Cream	7% w/v	3/3	3/3	3/3
Vagisil creme	7% w/v	3/3	3/3	3/3
VagiStat 1	7% w/v	3/3	3/3	3/3
Valacyclovir	2.5 mg/mL	3/3	3/3	3/3
Whole Blood	10% v/v	3/3	3/3	3/3
YeastGard Suppositories	7% w/v	3/3	3/3	3/3
* One out of three (1/3) initial replicates and two out of five (2/5) confirmation replicates were negative for HSV-2

CLINICAL AGREEMENT - Genital Swab Sample Type

Prospective Study

A total of 718 genital swab samples were prospectively collected from patients with signs and symptoms of genital herpes simplex virus (HSV) infection from 6 geographically diverse locations. Of the 718 samples collected. 9 samples were removed from the analysis because they were either not tested or had invalid results on the 3 assays (Simplexa™ HSV 1 & 2 Direct, Culture or bi-directional sequencing). Of the 709 remaining samples, 13 samples were removed from the analysis because they were not tested on the tests included in the comparator method sufficient to generate a final comparator result. A total of 696 samples were used for the analysis. The clinical performance of the Simplexa HSV 1 & HSV 2 assay was evaluated by comparing the positive and negative percent agreement to a composite comparator algorithm consisting of; culture, bi-directional sequencing and an FDA cleared NAAT. A positive result for HSV-1 and/or HSV-2 was determined by a positive test result in either the culture or the bi-directional sequencing. If both the culture and the bi-directional sequencing yielded positive results but disagreed in the differentiation of HSV-1 versus HSV-2, the results of the FDA cleared NAAT were used and a 2 out of 3 rule was followed to determine the type of the virus (e.g. if two of the methods were positive for HSV-1, the final comparator result was HSV-1 positive). All sites collected the genital swab samples on the Simplexa™ HSV-1 and HSV-2 Direct and sent samples to a central lab for culture testing. For culture, each sample was tested for HSV- 2 first and if positive for HSV-2 no further testing was performed. Samples that were HSV-2 culture negative were further tested for HSV-1 culture positivity. Dual positives could not be identified in the culture assay.

The available retained samples were sent to Focus Diagnostics and tested in a validated bi-directional sequencing assay. Of the 24 discordant samples that were positive for HSV-2 by the culture method but positive for HSV-1 by the bi-directional sequencing assay, 18 samples had valid results and 1 sample had an invalid result when tested on an FDA cleared NAAT. There were 5 samples that were not tested on the FDA cleared NAAT due to insufficient volume and therefore 6 out of 24 samples were excluded from the

11

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510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 9 of 12

analysis. There were 2 samples that were positive for HSV-1 by the culture method but positive for HSV-2 by the bi-directional sequencing assay that were not tested on the FDA cleared NAAT for insufficient volume and therefore were excluded from analysis. Results for Simplexa™ HSV 1 & 2 Direct compared to the composite comparator algorithm are presented in the following tables.

Simplexa™ HSV 1 & 2 Direct Compared to Composite Comparator Result (HSV-1)

| Simplexa™ HSV 1 & 2 Direct

Results HSV-1	Composite Comparator Result (HSV-1)
	Detected	Not Detected	Total
Detected	111	10	121
Not Detected	3	560	563
Total	114	570	684

Sensitivity	97.4%(111/114)
95% CI: 92.5% to 99.1%	Specificity	98.2%(560/570)
95% CI: 96.8% to 99.0%
PPV	91.7%(111/121)
95% CI: 85.5% to 95.4%	NPV	99.5%(560/563)
95% CI: 98.4% to 99.8%

Simplexa™ HSV 1 & 2 Direct Compared to Composite Comparator Result (HSV-2)

Simplexa™ HSV 1 & 2 Direct	Composite Comparator Result (HSV-2)
Results HSV-2	Detected	Not Detected	Total
Detected	175	11	186
Not Detected	5	497	502
Total	180	508	688

Sensitivity	97.2%(175/180)
95% Cl: 93.7% to 98.8%	Specificity	97.8%(497/508)
95% CI: 96.2% to 98.8%
PPV	94.1%(175/186)
95% Cl: 89.7% to 96.7%	NPV	99.0%(497/502)
95% CI: 97.7% to 99.6%

12

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510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 10 of 12

Retrospective Study

A total of 28 genital swab samples (14 positive HSV-1 and 14 positive HSV-2) were retrospectively collected from male patients with signs and symptoms of genital herpes simplex virus (HSV) infection and contained preselected positive and negative samples were tested at Focus Diagnostics using the SimplexaTM HSV 1 & 2 Direct and a validated bi-directional sequencing assay.

HSV-1 Validated Bi-Directional Sequencing Assay Results

| Simplexa™ HSV 1 & 2 Direct

Results	Composite Comparator Result (HSV-1)
	Detected	Not Detected	Total
Detected	14	1	15
Not Detected	0	13	13
Total	14	14	28

PPA	100.0%(14/14)	NPA	92.9%(13/14)
	95% CI: 78.5% to 100.0%		95% CI: 68.5% to 98.7%
PPV	93.3%(14/15)	NPV	100.0%(13/13)
	95% CI: 70.2% to 98.8%		95% CI: 77.2% to 100.0%

HSV-2 Validated Bi-Directional Sequencing Assay Results

| Simplexa™ HSV 1 & 2 Direct

Results	Composite Comparator Result (HSV-2)
Results	Detected	Not Detected	Total
Detected	14	0	14
Not Detected	0	14	14
Total	14	14	28

PPA	100.0%(14/14)
95% CI: 78.5% to 100.0%	NPA	100.0%(14/14)
95% CI: 78.5% to 100.0%
PPV	100.0%(14/14)
95% CI: 78.5% to 100.0%	NPV	100.0%(14/14)
95% CI: 78.5% to 100.0%

13

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510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 11 of 12

EXPECTED VALUES - Genital Swab Sample Type

The observed expected values using the Simplexa™ HSV 1 & 2 Direct assay are presented below for patients 17 years of age or older from the prospective study population. The data is stratified by age, and gender and collection sites.

			Sample Demographics by Simplexa™ HSV 1 & 2 Direct Assay
Gender	Age Group		Simplexa™ HSV 1 & 2 Direct Result
		All	HSV-1 &
HSV-2 Not
Detected	HSV-1 Detected
& HSV-2 Not
Detected	HSV-1 Not
Detected & HSV-2
Detected	HSV-1 & HSV-2
Dual Positive	Not-Evaluable
Female	17 Years of
age to 21
Years of age	98	41.8% (41/98)	25.5% (25/98)	30.6% (30/98)	0.0% (0/98)	2.0% (2/98)
	More than 21
Years of age	503	54.3% (273/503)	16.9% (85/503)	25.8% (130/503)	0.8% (4/503)	2.2% (11/503)
	All	601	52.2% (314/601)	18.3% (110/601)	26.6% (160/601)	0.7% (4/601)	2.2% (13/601)
Male	17 Years of
age to 21
Years of age	14	42.9% (6/14)	21.4% (3/14)	35.7% (5/14)	0.0% (0/14)	0.0% (0/14)
	More than 21
Years of age	66	68.2% (45/66)	10.6% (7/66)	18.2% (12/66)	1.5% (1/66)	1.5% (1/66)
	All	80	63.8% (51/80)	12.5% (10/80)	21.3% (17/80)	1.3% (1/80)	1.3% (1/80)
All		681	53.6% (365/681)	17.6% (120/681)	26.0% (177/681)	0.7% (5/681)	0.7% (5/681)**

volume, daily PC/NTC failure,

** Excluded from this table are 35 samples from 17 years of age and 2 samples where the age of the patient was not t available.

Hypothetical Prevalence	HSV-1		HSV-2
	PPV*	NPV**	PPV*	NPV**
1.0%	35.3%	100.0%	30.9%	100.0%
2.0%	52.5%	99.9%	47.4%	99.9%
3.0%	62.6%	99.9%	57.7%	99.9%
5.0%	74.0%	99.9%	69.9%	99.8%
10.0%	85.7%	99.7%	83.1%	99.7%
15.0%	90.5%	99.5%	88.6%	99.5%
20.0%	93.1%	99.3%	91.7%	99.3%
30.0%	95.9%	98.9%	95.0%	98.8%
50.0%	98.2%	97.4%	97.8%	97.2%

*Positive Predictive Value (PPV) was calculated using:

(Sensitivity x Prevalence)/(Sensitivity x Prevalence + [1 - Specificity] x [1 - Prevalence]).

**Negative Predictive Value (NPV ) was calculated using:

(Specificity x [1 - Prevalence])/([1 - Sensitivity] x Prevalence + Specificity x [1 - Prevalence]).

14

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510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 August 20, 2015 Page 12 of 12

FRESH VS FROZEN STUDY

Storage conditions were validated using the following transport media types BD VTM, M4, M4RT, M5, M6, and UTM by spiking media with organism at concentrations ranging from 3 times LoD to 50 times LoD and at different storage temperatures and durations.

Samples should be transported on ice and stored at 2 to 8°C for up to 7 days post collection. If there is a greater than 7 day delay before processing of the sample, store the sample at -70° C.

CONCLUSION

The conclusions drawn from the nonclinical tests demonstrate the device is as safe and effective as the legally marketed device identified above.