Simplexa HSV 1 & 2 Direct, Simplexa HSV 1 & 2 Positive Control Pack

K173798 · Diasorin Molecular, LLC · PGI · Mar 14, 2018 · Microbiology

Device Facts

• Record ID: K173798
• Device Name: Simplexa HSV 1 & 2 Direct, Simplexa HSV 1 & 2 Positive Control Pack
• Applicant: Diasorin Molecular, LLC
• Product Code: PGI · Microbiology
• Decision Date: Mar 14, 2018
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3309
• Device Class: Class 2

Intended Use

The DiaSorin Molecular Simplexa™ HSV 1 & 2 Direct assay is intended for use on the LIAISON® MDX instrument for the qualitative detection and differentiation of herpes simplex virus (HSV-1 and HSV-2) DNA present in mucocutaneous and cutaneous lesion swabs from patients with signs and symptoms of HSV-1 or HSV-2 infection. This test is an aid in the differential diagnosis of HSV-1 and HSV-2 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. 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 Story

Simplexa HSV 1 & 2 Direct is a real-time PCR assay for qualitative detection and differentiation of HSV-1 and HSV-2 DNA from unprocessed cutaneous and mucocutaneous lesion swabs. Device eliminates nucleic acid extraction; uses bi-functional fluorescent probe-primers targeting conserved regions of HSV-1 and HSV-2 DNA polymerase genes. System includes assay reagents, LIAISON MDX instrument, LIAISON MDX Studio Software, and Direct Amplification Disc. Operated by laboratory professionals in clinical settings. Input: lesion swab specimens. Output: qualitative detection of HSV-1, HSV-2, and internal control. Internal control monitors for PCR failure/inhibition. Results aid clinicians in differential diagnosis of HSV infections. Benefits include rapid, direct testing without extraction, facilitating timely clinical management.

Clinical Evidence

Clinical performance evaluated via prospective and retrospective studies comparing Simplexa HSV 1 & 2 Direct to a composite reference method (culture, bi-directional sequencing, and FDA-cleared NAAT). Prospective study (n=514) and retrospective study (n=365) data combined. For cutaneous swabs, HSV-1 PPA 100% (30/30), NPA 96.3% (182/189); HSV-2 PPA 97.0% (32/33), NPA 97.9% (182/186). For mucocutaneous swabs, HSV-1 PPA 98.2% (162/165), NPA 97.5% (703/721); HSV-2 PPA 99.5% (193/194), NPA 96.7% (669/692). Analytical studies confirmed no cross-reactivity or interference with 71 microorganisms and 24 substances.

Technological Characteristics

Real-time PCR assay; targets HSV-1 and HSV-2 DNA polymerase genes. Uses bi-functional fluorescent probe-primers (HSV-1: CFR610; HSV-2: FAM; Internal Control: Q670). Direct amplification (no extraction). Platform: LIAISON MDX instrument with Direct Amplification Disc. Software: LIAISON MDX Studio Software. Connectivity: Standalone instrument. Sterilization: Not applicable (reagents).

Indications for Use

Indicated for qualitative detection and differentiation of HSV-1 and HSV-2 DNA in mucocutaneous and cutaneous lesion swabs from symptomatic patients. For professional use only; prescription use only.

Regulatory Classification

Identification

A herpes virus nucleic acid-based cutaneous and mucocutaneous lesion panel is a qualitative in vitro diagnostic device intended for the simultaneous detection and differentiation of different herpes viruses in cutaneous and mucocutaneous lesion samples from symptomatic patients suspected of Herpetic infections. Negative results do not preclude infection and should not be used as the sole basis for treatment or other patient management decisions. The assay is not intended for use in cerebrospinal fluid samples.

Special Controls

*Classification.* Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include detailed documentation for the device description, including the device components, ancillary reagents required but not provided, and a detailed explanation of the methodology including primer design and selection. (2) Premarket notification submissions must include detailed documentation from the following analytical and clinical performance studies: Analytical sensitivity (Limit of Detection), reactivity, inclusivity, precision, reproducibility, interference, cross reactivity, carry-over, and cross contamination. (3) Premarket notification submissions must include detailed documentation of a clinical study using lesion samples in which Herpes Simplex Virus 1, Herpes Simplex Virus 2, or Varicella Zoster Virus DNA detection was requested. The study must compare the device performance to an appropriate well established reference method. (4) A detailed explanation of the interpretation of results and acceptance criteria must be included in the device's 21 CFR 809.10(b)(9) compliant labeling. (5) The device labeling must include a limitation statement that reads: “The device is not intended for use with cerebrospinal fluid or to aid in the diagnosis of HSV or VZV infections of the central nervous system (CNS).” (6) Premarket notification submissions must include quality assurance protocols and a detailed documentation for device software, including, but not limited to, standalone software applications and hardware-based devices that incorporate software. (7) The risk management activities performed as part of the manufacturer's 21 CFR 820.10(c) design and development activities must document an appropriate end user device training program that will be offered as part of efforts to mitigate the risk of failure to correctly operate the instrument.

Predicate Devices

• Lyra® Direct HSV 1 + 2/VZV Assay (K133448)

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo in blue, with the words "U.S. FOOD & DRUG" stacked on top of the word "ADMINISTRATION".

March 14, 2018

DiaSorin Molecular LLC Sharon Young Senior Regulatory Affairs Specialist 11331 Valley View Street Cypress, California 90630

Re: K173798

Trade/Device Name: Simplexa HSV 1 & 2 Direct, Simplexa HSV 1 & 2 Positive Control Pack Regulation Number: 21 CFR 866.3309 Regulation Name: Herpes Virus Nucleic Acid-Based Cutaneous and Mucocutaneous Lesion Panel Regulatory Class: Class II Product Code: PGI Dated: December 12, 2017 Received: December 14, 2017

Dear Sharon 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 and Part 809); medical device 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

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

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.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

# Steven R. Gitterman -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

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### Indications for Use

510(k) Number (if known)

Device Name

Simplexa™ HSV 1 & 2 Direct MOL2150 and Simplexa™ HSV 1 & 2 Positive Control Pack MOL2160

Indications for Use (Describe)

Simplexa™ HSV 1 & 2 Direct MOL2150

The DiaSorin Molecular Simplexa™ HSV 1 & 2 Direct assay is intended for use on the LIAISON® MDX instrument for the qualitative detection and differentiation of herpes simplex virus (HSV-2) DNA present in mucocutaneous and cutaneous lesion swabs from patients with signs and symptoms of HSV-1 or HSV-2 infection. This test is an aid in the differential diagnosis of HSV-1 and HSV-2 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 MOL2160

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.

Type of Use (Select one or both, as applicable)

| <span style="font-size: 16px;">☑</span> Prescription Use (Part 21 CFR 801 Subpart D) |
|--------------------------------------------------------------------------------------|
| <span style="font-size: 16px;">☐</span> 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 DiaSorin Molecular. The logo features a stylized DNA double helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in a dark blue, sans-serif font, with the word "Molecular" underneath in a lighter green color. The overall design is clean and modern, suggesting a company focused on molecular diagnostics or related fields.

510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 March 9. 2018 Page 1 of 20

| Applicant | DiaSorin Molecular LLC.<br>11331 Valley View Street<br>Cypress, California 90630<br>USA |
|-------------------------------------------|-------------------------------------------------------------------------------------------------------|
| Establishment Registration No. | 2023365 |
| Contact Person | Sharon Young<br>tel [phone]<br>fax [phone]<br>[e-mail] |
| Summary Date | March 9, 2018 |
| Proprietary Name | Simplexa™ HSV 1 & 2 Direct MOL2150 and Simplexa™ HSV 1 & 2<br>Positive Control Pack MOL2160 |
| Generic Name | HSV 1 & 2 nucleic acid |
| Classification Regulation<br>Product Code | PGI |
| Regulation Number | 866.3309 |
| Regulation name | Herpes Virus (VZV, HSV1, HSV2), DNA Detection Assay for<br>Cutaneous and Mucocutaneous Lesion Samples |
| Predicate Devices | Lyra® Direct HSV 1 + 2/VZV Assay K133448 |

### Intended Use

Simplexa™ HSV 1 & 2 Direct

The DiaSorin Molecular Simplexa™ HSV 1 & 2 Direct assay is intended for use on the LIAISON® MDX instrument for the qualitative detection and differentiation of herpes simplex virus (HSV-1 and HSV-2) DNA present in mucocutaneous and cutaneous lesion swabs from patients with signs and symptoms of HSV-1 or HSV-2 infection. This test is an aid in the differential diagnosis of HSV-1 and HSV-2 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 cutaneous and mucocutaneous lesion swab specimens without nucleic acid extraction. The system consists of the Simplexa™ HSV 1 & 2 Direct assay, the LIAISON® MDX (with LIAISON® MDX 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 DiaSorin Molecular. The logo consists of a green and blue DNA helix on the left, followed by the text "DiaSorin" in blue and "Molecular" in green. The text is aligned to the right of the DNA helix.

### 510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 March 9, 2018 Page 2 of 20

## Simplexa™ HSV 1 & 2 Direct REF MOL2150

| Component Name | REF | EC SYMBOL<br>ON LABEL | Abbreviated<br>Name | Cap<br>Color | Number<br>of Vials | Reactions<br>per<br>Vial/Kit | Volume<br>per Vial | |
|--------------------------------------------|---------|-----------------------|---------------------|--------------|--------------------|------------------------------|--------------------|-------|
| Simplexa™ HSV 1 & 2 Direct<br>Reaction Mix | MOL2151 | REAG | C | RM | Brown | 24 | 1/24 | 50 μL |

#### Component Description

| Kit Component | Contents | | | | | | | | | | | | | | | | | | | | |
|----------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|
| Simplexa™ HSV 1<br>& 2 Direct Reaction<br>Mix (RM) | DNA polymerase, buffer, dNTPs, template DNA (Internal Control) dye-labeled fluorescent<br>probe-primers specific for detection of HSV-1 and/or HSV-2 and for the DNA Internal<br>Control. | | | | | | | | | | | | | | | | | | | | |
| | Target Probe<br>Fluorophore<br>(Dye) Excitation<br>(nm) Emission<br>(nm) Targeted Gene HSV-1 CFR610 590 610 HSV-1 DNA<br>polymerase HSV-2 FAM 495 520 HSV-2 DNA<br>polymerase DNA Internal<br>Control Q670 644 670 NA | | | | | | | | | | | | | | | | | | | | |
| Simplexa™ HSV 1<br>& 2 Kit Barcode<br>Card | Assay specific parameters. | | | | | | | | | | | | | | | | | | | | |

### Simplexa™ HSV 1 & 2 Positive Control Pack REF MOL2160

| Component Name | REF | Description | Cap Color | Number<br>of Vials | Reactions per<br>Vial/Kit | Volume per<br>Vial |
|------------------------------------------------|---------|------------------------------------|-----------|--------------------|---------------------------|--------------------|
| Simplexa™ HSV 1 & 2<br>Direct Positive Control | MOL2161 | Inactivated HSV<br>1 and 2 viruses | Red | 10 | 1/10 | 100 µL |

### Direct Amplification Disc kit REF MOL1455

| Component Name | REF | Description | Number of<br>Discs | Reactions<br>per Disc/Kit |
|---------------------------|---------|------------------------------------------------------------------------------------------------|--------------------|---------------------------|
| Direct Amplification Disc | MOL1452 | Direct Amplification Disc for the<br>processing of up to 8 individual controls<br>or specimens | 3 | 8/24 |

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Image /page/5/Picture/0 description: The image contains the logo for DiaSorin Molecular. The logo consists of a green and blue DNA helix graphic on the left, followed by the text "DiaSorin" in dark blue. Below "DiaSorin" is the word "Molecular" in green.

### 510(k) Summary

March 9, 2018 Page 3 of 20

### Predicate Device Information

| Table of Similarities | | |
|-------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | Predicate | Device |
| Name | Lyra® Direct HSV 1 + 2/VZV Assay<br>K133448 | Simplexa™ HSV<br>1 & 2 Direct |
| Intended<br>Use | The Lyra® Direct HSV 1 + 2/VZV Assay is an<br>in vitro multiplex Real-Time PCR test for<br>qualitative detection and differentiation of<br>herpes simplex virus type 1, herpes simplex<br>virus type 2, and varicella-zoster virus DNA<br>isolated and purified from cutaneous or<br>mucocutaneous lesion samples obtained from<br>symptomatic patients suspected of active<br>herpes simplex virus 1, herpes simplex virus 2<br>and/or varicella-zoster infection. The Lyra®<br>Direct HSV 1 + 2/VZV Assay is intended to aid<br>in the diagnosis of herpes simplex virus 1,<br>herpes simplex virus 2 and varicella-zoster<br>virus active cutaneous or mucocutaneous<br>infections. Negative results do not preclude<br>herpes simplex virus 1, herpes simplex virus 2<br>and varicella-zoster virus infections and should<br>not be used as the sole basis for diagnosis,<br>treatment or other management decisions. The<br>Lyra® Direct HSV 1 + 2/VZV Assay is not<br>intended for use with cerebrospinal fluid or to<br>aid in the diagnosis of HSV or VZV infections<br>of the central nervous system (CNS). The<br>Lyra® Direct HSV 1 + 2/VZV Assay is not<br>intended for use in prenatal screening. The<br>device is not intended for point-of-care use. | The DiaSorin Molecular Simplexa™ HSV 1 & 2<br>Direct assay is intended for use on the LIAISON®<br>MDX instrument for the qualitative detection and<br>differentiation of herpes simplex virus (HSV-1 and<br>HSV-2) DNA present in mucocutaneous and<br>cutaneous lesion swabs from patients with signs<br>and symptoms of HSV-1 or HSV-2 infection. This<br>test is an aid in the differential diagnosis of HSV-1<br>and HSV-2 infections.<br><br>The assay is not intended for use as a screening<br>test for the presence of HSV-1 and HSV-2 in<br>blood or blood products. The assay is for<br>professional use only<br><br>Simplexa™ HSV 1 & 2 Positive Control Pack<br>The Simplexa™ HSV 1 & 2 Positive Control Pack<br>is intended to be used as a control with the<br>Simplexa™ HSV 1 & 2 Direct kit.<br>This control is not intended for use with other<br>assays or systems. |
| Sample<br>Types | Cutaneous or mucocutaneous lesion samples | Same |
| Extraction<br>Methods | None | Same |
| Assay<br>Methodology | PCR-based system for detecting the presence<br>or absence of viral DNA in clinical specimens. | Same |
| Detection<br>Techniques | Multiplex assay using different reporter dyes<br>for each target. | Same |
| Table of Differences | | |
| Name | Predicate | Candidate |
| | Lyra® Direct HSV 1 + 2/VZV Assay<br>K133448 | Simplexa™ HSV<br>1 & 2 Direct |
| Assay<br>Targets | HSV-1: glycoprotein G, HSV-2: glycoprotein G,<br>VZV: ORF6: DNA-helicase primase. | Well conserved region of the polymerase genes<br>for HSV-1 and HSV-2. |

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Image /page/6/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a green and blue DNA helix on the left, with the words "DiaSorin" in dark blue on the top right. Below "DiaSorin" is the word "Molecular" in green.

### 510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 March 9. 2018 Page 4 of 20

### ANALYTICAL STUDIES

Results of the analytical studies performed were combined with analytical results from K150962.

### REPRODUCIBILITY

Reproducibility studies were not conducted again for the purposes of this submission and the results from original submission K150962 are shown below.

Reproducibility for the Simplexa™ HSV 1 & 2 Direct assay was evaluated. Three investigative sites assessed the device's inter-site, inter-day and interlintra-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 negative approximately 95% of the time. The assays were performed in triplicate on 5 different days. Each site had 2 operators; each operator assayed the entire sample panel and positive control once per day, for a total of 2 sets of data per day. Combined results for all sites are presented in the tables below.

| | | Site - 1 | | | Site - 2 | | | Site - 3 | | | Total % Agreement With Expected Results | 95% CI | |
|----------------------------------------------------------------------------------------------------------------------|-----------------------------|-----------------------------------------------------------------------------------|---------------------|-----------------------|-----------------------------------------------------------------------------------|---------------------|-----------------------|-----------------------------------------------------------------------------------|---------------------|-----------------------|-----------------------------------------------------------------------------------------|-------------------|--|
| | 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 | | | |
| | 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 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-<br>1<br>Result | 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. | | | | | | | | | | | | | |
| | | | Site - 1 | | | Site - 2 | | | Site - 3 | | | | |
| | 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 | |
| HSV-<br>2<br>Result | HSV-1 Low<br>Positive | 100.0%<br>(30/30) | NA | NA | 100.0%<br>(30/30) | NA | NA | 96.7%<br>(29/30)b | 41.1 | 0.0 | 98.9%<br>(89/90)b | 94.0 to<br>99.8% | |
| | HSV-1<br>Medium<br>Positive | 100.0%<br>(30/30) | NA | NA | 100.0%<br>(30/30) | NA | NA | 100.0%<br>(30/30)b | NA | NA | 100.0%<br>(90/90) | 95.9 to<br>100.0% | |
| | HSV-2 Low<br>Positive | 100.0%<br>(30/30) | 37.4 | 2.9 | 90.0%<br>(27/30) | 37.5 | 3.5 | 93.3%<br>(28/30) | 37.1 | 2.8 | 94.4%<br>(85/90) | 87.6 to<br>97.6% | |
| | HSV-2<br>Medium<br>Positive | 100.0%<br>(30/30) | 35.5 | 1.9 | 100.0%<br>(30/30) | 35.6 | 2.0 | 100.0%<br>(30/30) | 35.3 | 1.6 | 100.0%<br>(90/90) | 95.9 to<br>100.0% | |
| | High Negative | 96.7%<br>(29/30)b | 39.5 | 0.0 | 86.7%<br>(26/30)b | 38.6 | 2.9 | 100.0%<br>(30/30)b | NA | NA | 94.4%<br>(85/90)b | 87.6 to<br>97.6% | |
| | Positive<br>Control | 100.0%<br>(30/30) | 30.2 | 1.3 | 100.0%<br>(30/30) | 30.1 | 0.6 | 100.0%<br>(29/29) | 29.9 | 1.2 | 100.0%<br>(89/89) | 95.9 to<br>100.0% | |
| | Total<br>Agreement | 99.4% (179/180) | | | 96.1% (173/180) | | | 98.9% (176/179) | | | 98.0%<br>(528/539) | 96.4 to<br>98.9% | |
| b) Expected Results of HSV-1 Low Positive, HSV-1 Medium Positive and High Negative samples are "Negative" for HSV-2. | | | | | | | | | | | | | |
| | | | Site - 1 | | | Site - 2 | | | Site - 3 | | | | |
| | 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<br>IC<br>Result | HSV-1 Low<br>Positive | 100.0%<br>(30/30) | 29.6 | 0.7 | 100.0%<br>(30/30) | 29.8 | 1.2 | 100.0%<br>(30/30) | 29.7 | 1.0 | 100.0%<br>(90/90) | 95.9 to<br>100.0% | |
| | HSV-1<br>Medium<br>Positive | 100.0%<br>(30/30) | 29.6 | 0.8 | 100.0%<br>(30/30) | 29.8 | 1.4 | 100.0%<br>(30/30) | 29.7 | 0.9 | 100.0%<br>(90/90) | 95.9 to<br>100.0% | |
| | HSV-2 Low<br>Positive | 100.0%<br>(30/30) | 29.6 | 0.8 | 100.0%<br>(30/30) | 29.8 | 1.2 | 100.0%<br>(30/30) | 29.7 | 1.0 | 100.0%<br>(90/90) | 95.9 to<br>100.0% | |
| | HSV-2<br>Medium<br>Positive | 100.0%<br>(30/30) | 29.5 | 0.6 | 100.0%<br>(30/30) | 29.7 | 1.4 | 100.0%<br>(30/30) | 29.8 | 1.4 | 100.0%<br>(90/90) | 95.9 to<br>100.0% | |
| | High<br>Negative | 100.0%<br>(30/30) | 29.6 | 0.6 | 100.0%<br>(30/30) | 29.8 | 1.2 | 100.0%<br>(30/30) | 29.7 | 1.0 | 100.0%<br>(90/90) | 95.9 to<br>100.0% | |
| | Positive<br>Control | 100.0%<br>(30/30) | 29.5 | 0.5 | 100.0%<br>(30/30) | 29.7 | 1.4 | 100.0%<br>(29/29) | 29.7 | 0.9 | 100.0%<br>(89/89) | 95.9 to<br>100.0% | |
| | Total<br>Agreement | 100.0% (180/180) | | | 100.0% (180/180) | | 100.0% (179/179) | | | 100.0%<br>(539/539) | 96.4 to<br>98.9% | | |

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Image /page/7/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a green and blue DNA helix graphic on the left. To the right of the graphic is the text "DiaSorin" in blue, with the word "Molecular" underneath in green.

## 510(k) Summary

March 9, 2018
Page 5 of 20

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Image /page/8/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a stylized DNA double helix on the left, with the words "DiaSorin" in a dark blue sans-serif font to the right of the helix. Below "DiaSorin" is the word "Molecular" in a green sans-serif font. The DNA helix is colored with a gradient that transitions from green at the top to blue at the bottom.

510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2150 Simplexa™ HSV 1 & 2 Positive Control Pack Catalog No. MOL2160 March 9. 2018 Page 6 of 20

### ANALYTICAL SENSITIVITY/LIMIT OF DETECTION

Analytical Sensitivity/Limit of Detection studies were not conducted again for the purposes of this submission and the results from original submission K150962 are shown below.

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 cutaneous and mucocutaneous swab matrix. LoD was determined to be the lowest concentration that could be detected positive > 95% of the time.

| Virus Strain | LoD Concentration<br>(TCID50/mL) | Qualitative Results<br>(#Detected/#Total) | Mean Ct ± SD<br>(from Detected<br>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 |

### ANALYTICAL REACTIVITY / CROSS REACTIVITY

### Analytical Reactivity Cutaneous and Mucocutaneous Swab Sample Type

Analytical Reactivity studies were not conducted again for the purposes of this submission and the results from original submission K150962 are shown below.

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 cutaneous swab matrix containing male and female cutaneous and mucocutaneous 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<br>[TCID50/mL] | Qualitative Result<br>(#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) Cutaneous and Mucocutaneous 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 cutaneous or mucocutaneous swabs. A total of 71 potential cross-reactants were spiked into negative cutaneous and mucocutaneous swab matrix and assayed in triplicate. No cross-reactivity was observed.

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Image /page/9/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA double helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in a dark blue, sans-serif font, with the word "Molecular" underneath in a lighter green color.

### 510(k) Summary

March 9, 2018
Page 7 of 20

| No. | Microorganism | Tested Concentration | Qualitative Result<br>(#Detected/#Total) | |
|-----|--------------------------------------------------|-------------------------------|------------------------------------------|-------|
| | | | HSV-1 | HSV-2 |
| 1 | Baseline | N/A | 35/35 | 35/35 |
| 2 | Acinetobacter calcoaceticus | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 3 | Acinetobacter Iwoffii | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 4 | Bacteroides fragilis | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 5 | Bacteroides ureolyticus** | N/A | N/A | N/A |
| 6 | Bordetella bronchiseptica | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 7 | Bordetella pertussis | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 8 | Candida albicans | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 9 | Candida glabrata | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 10 | Candida guilliermondii | 1.00 x 106 CFU/mL | 0/3 | 0/3 |
| 11 | Candida krusei | 1.00 x 106 CFU/mL | 0/3 | 0/3 |
| 12 | Candida lusitaniae | 1.00 x 106 CFU/mL | 0/3 | 0/3 |
| 13 | Candida parapsilosis | 1.00 x 106 CFU/mL | 0/3 | 0/3 |
| 14 | Candida tropicalis | 1.00 x 106 CFU/mL | 0/3 | 0/3 |
| 15 | Chlamydophila pneumoniae | 1.00 x 106 IFU/mL | 0/3 | 0/3 |
| 16 | Chlamydia trachomatis | 1.00 X 106 IFU/mL | 0/3 | 0/3 |
| 17 | Clostridium sordellii | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 18 | Clostridium perfringens | 1.00 x 106 CFU/mL | 0/3 | 0/3 |
| 19 | Corynebacterium genitalium | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 20 | Coronavirus (HCoV OC43) | 1.00 x 105 TCID50/mL | 0/3 | 0/3 |
| 21 | Corynebacterium diphtheriae | 1.00 x 106 CFU/mL | 0/3 | 0/3 |
| 22 | Coxsackievirus B (CVB-1) | 1.00 x 105 TCID50/mL | 0/3 | 0/3 |
| 23 | Cytomegalovirus | 1.00 X 105 TCID50/mL | 0/3 | 0/3 |
| 24 | Enterobacter cloacae | 1.00 x 106 CFU/mL | 0/3 | 0/3 |
| 25 | Enterococcus faecium | 1.00 x 106 CFU/mL | 0/3 | 0/3 |
| 26 | Enterococcus faecalis vanB | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 27 | Enterovirus 70 | 1.00 x 105 TCID50/mL | 0/3 | 0/3 |
| 28 | Enterovirus 71 | 1.00 X 105 TCID50/mL | 0/3 | 0/3 |
| 29 | Epstein Barr Virus (B95-8) | 1.00 X 105 copies/mL | 0/3 | 0/3 |
| 30 | Escherichia coli O157H7 | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 31 | Fusobacterium nucleatum | 1.00 x 106 CFU/mL | 0/3 | 0/3 |
| 32 | Gardnerella vaginalis | 1.00 X 106 CFU/mL | 0/3 | 0/3 |
| 33 | Haemophilus ducreyi** | N/A | N/A | N/A |
| 34 | Haemophilus influenzae (Type A) | 1.00 x 106 CFU/mL | 0/3 | 0/3 |
| 35 | Hepatitis B | 1.00 X 105 IU/mL | 0/3 | 0/3 |
| 36 | Hepatitis C | 1.00 X 105 IU/mL | 0/3 | 0/3 |
| 37 | HHV-6 (Z29 Strain) | 1.00 X 105 TCID50/mL | 0/3 | 0/3 |
| 38 | HHV-7 SB | 1.00 X 105 TCID50/mL | 0/3 | 0/3 |
| No. | Microorganism | Tested Concentration | Qualitative Result<br>(#Detected/#Total) | |
| | | | HSV-1 | HSV-2 |
| 39 | HIV-1 IIIB | $1.00 X 10^5$ copies/mL | 0/3 | 0/3 |
| 40 | HIV-2 NIHZ* | Not Available | 0/3 | 0/3 |
| 41 | HPV18 Recombinant | $1.00 X 10^5$ PFU/mL | 0/3 | 0/3 |
| 42 | Human metapneumovirus | $1.00 X 10^5$ TCID50/mL | 0/3 | 0/3 |
| 43 | Lactobacillus acidophilus | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 44 | Legionella pneumophila | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 45 | Mobiluncus mulieris | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 46 | Moraxella catarrhalis | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 47 | Mycoplasma genitalium** | N/A | N/A | N/A |
| 48 | Mycoplasma hominis | $1.00 X 10^6$ CCU/mL | 0/3 | 0/3 |
| 49 | Mycoplasma orale** | N/A | N/A | N/A |
| 50 | Mycoplasma pneumoniae | $1.00 X 10^6$ CCU/mL | 0/3 | 0/3 |
| 51 | Mycoplasma salivarium** | N/A | N/A | N/A |
| 52 | Neisseria gonorrhoeae | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 53 | Neisseria meningitides | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 54 | Prevotella melaninogenica | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 55 | Proteus vulgaris | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 56 | Respiratory syncytial virus A | $1.00 X 10^5$ TCID50/mL | 0/3 | 0/3 |
| 57 | Respiratory syncytial virus B | $1.00 X 10^5$ TCID50/mL | 0/3 | 0/3 |
| 58 | Rubella | $1.00 X 10^5$ TCID50/mL | 0/3 | 0/3 |
| 59 | Salmonella enteritidis** | N/A | N/A | N/A |
| 60 | Salmonella typhimurium | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 61 | Staphylococcus aureus (MRSA), ATCC<br>700699 | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 62 | Staphylococcus epidermidis (MRSE),<br>ATCC 29887 | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 63 | Streptococcus mutans | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 64 | Streptococcus salivarius | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 65 | Staphylococcus saprophyticus | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 66 | Streptococcus mitis | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 67 | Streptococcus pyogenes, M1 | $1.00 X 10^6$ CFU/mL | 0/3 | 0/3 |
| 68 | Toxoplasma gondii | $1.00 X 10^6$ tachyzooites/mL | 0/3 | 0/3 |
| 69 | Treponema pallidum** | N/A | N/A | N/A |
| 70 | Trichomonas vaginalis | $1.00 X 10^6$ trophozoites/ml | 0/3 | 0/3 |
| 71 | Ureaplasma urealyticum | $1.00 X 10^6$ CCU/mL | 0/3 | 0/3 |
| 72 | VZV | $1.00 X 10^5$ copies/mL | 0/3 | 0/3 |

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Image /page/10/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA double helix in shades of green and blue on the left. To the right of the DNA helix are the words "DiaSorin" in a dark blue, sans-serif font, stacked above the word "Molecular" in a lighter green, sans-serif font.

### 510(k) Summary

Simplexa™ HSV 1 & 2 Direct Catalog No. MOL2…