Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The device uses "on-board method-specific algorithms" to interpret fluorescent signals, which is standard for many diagnostic instruments and does not indicate the use of AI or ML. The document does not mention AI, ML, or related terms.

Therapeutic

No
The device is an in vitro diagnostic test intended to aid in the diagnosis of infections by detecting viral DNA, not to treat them.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states, "The Solana® HSV 1+2/VZV Assay is an in vitro diagnostic test... The Solana® HSV 1+2/VZV Assay is intended to aid in the diagnosis of herpes simplex virus 1, herpes simplex virus 2 and varicella-zoster virus active cutaneous or mucocutaneous infections."

SaMD (Software as a Medical Device)

No

The device is an in vitro diagnostic test that includes reagents and is intended for use with a specific hardware instrument (Solana® instrument) for amplification and detection. It is not solely software.

IVD (In Vitro Diagnostic)

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

The "Intended Use / Indications for Use" section explicitly states: "The Solana® HSV 1+2/VZV Assay is an in vitro diagnostic test..."

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Solana® HSV 1+2/VZV Assay is an in vitro diagnostic test, using isothermal amplification technology (helicase-dependent amplification, HDA), for the qualitative detection and differentiation of herpes simplex virus type 1, herpes simplex virus type 2, and varicella-zoster virus DNA isolated and purified from cutaneous lesion samples obtained from symptomatic patients suspected of active herpes simplex virus 1, herpes simplex virus 2 and/or varicella-zoster infection. The Solana® HSV 1+2/VZV Assay is intended to aid in the diagnosis of herpes virus 1. herpes simplex virus 2 and varicella-zoster virus active cutaneous infections. Negative results do not preclude herpes simplex virus 1, herpes simplex virus 2 and varicella-zoster virus infections and should not be used as the sole basis for diagnosis, treatment or other management decisions. The Solana® HSV 1+2/VZV Assay is intended for use only with the Solana® instrument.

Warning: The Solana® HSV 1+2/VZV Assay 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). The Solana® HSV 1+2/VZV Assay is not intended for use in prenatal screening.

Product codes

PGI

Device Description

The Solana® HSV 1+2/VZV Assay amplifies and detects viral DNA isolated from cutaneous or mucocutaneous lesion samples obtained from symptomatic patients suspected of active herpes simplex virus 1, herpes simplex virus 2 and/or varicella-zoster infection.

The assay consists of two (2) major steps: 1) specimen preparation, and 2) amplification and detection of target sequence specific to HSV-1, HSV-2 and/or VZV using isothermal Helicase-Dependent Amplification (HDA) in the presence of target-specific fluorescence probe.

Patient specimen is transferred to a Process Tube, subjected to heat treatment at 95°C for 5 minutes and mixed and vortexed. The processed sample is transferred to a Reaction Tube and mixed. The Reaction Tube contains lyophilized HDA reagents, dNTPs, primers and probes. Once rehydrated with the diluted sample, the Reaction Tube is placed in Solana for amplification and detection of specific target sequence. In Solana, the target sequences are amplified by HSV-1, HSV-2 and/or VZV specific primers and detected by HSV-1, HSV-2 and/or VZV specific fluorescence probes included in the Reaction Tube. A competitive process control (PRC) is included in the Process Tube to monitor sample processing, inhibitory substances in clinical samples, reagent failure or device failure. The PRC target is amplified by specific primers and detected by a PRC specific fluorescence probe.

The target and PRC probes are labeled with a quencher on one end and a fluorophore on the other end. In addition, the target and PRC probes carry a ribonucleic acid. Upon annealing to HSV-1, HSV-2, VZV or PRC amplicons, the fluorescence probes are cleaved by RNaseH2 and the fluorescence signal increases due to physical separation of fluorophore from quencher. Solana measures and interprets the fluorescent signal, using on-board method-specific algorithms. Solana will then report the test results to the user on its display screen, and the results can be printed via an attached printer.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

cutaneous or mucocutaneous lesion samples

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

Performance characteristics of the Solana " HSV 1+2/VZV Assay were established during a prospective study between February and April 2016. One thousand sixty-two (1062) fresh lesion specimens, collected for herpes simplex/varicella-zoster identification, have been included in this study at three sites across the United States. A single specimen was collected per patient. The specimens were processed and tested with Solana " HSV 1+2/VZV Assay on the Solana "instrument at the sites. Testing of the comparator methods (culture with DFA) were performed at one central location.

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

Clinical Sensitivity:
Performance characteristics of the Solana " HSV 1+2/VZV Assay were established during a prospective study between February and April 2016. One thousand sixty-two (1062) fresh lesion specimens, collected for herpes simplex/varicella-zoster identification, have been included in this study at three sites across the United States. A single specimen was collected per patient. The specimens were processed and tested with Solana " HSV 1+2/VZV Assay on the Solana "instrument at the sites. Testing of the comparator methods (culture with DFA) were performed at one central location.

Combined Data
Cutaneous Lesions (N=249 for HSV-1, N=275 for HSV-2, N=222 for VZV):

HSV-1 (Comparator: ELVIS® HSV ID and D3 Typing Test): Sensitivity 100% (22/22); Specificity 97.8% (222/227).
HSV-2 (Comparator: ELVIS® HSV ID and D3 Typing Test): Sensitivity 92.3% (24/26); Specificity 94.4% (235/249).
VZV (Comparator: DSFA and Culture with DFA): Sensitivity 100% (22/22); Specificity 96.5% (193/200).

Mucocutaneous Lesions (N=501 for HSV-1, N=610 for HSV-2, N=372 for VZV):

HSV-1 (Comparator: ELVIS® HSV ID and D3 Typing Test): Sensitivity 100% (113/113); Specificity 96.4% (374/388).
HSV-2 (Comparator: ELVIS® HSV ID and D3 Typing Test): Sensitivity 99.1% (108/109); Specificity 97.2% (487/501).
VZV (Comparator: DSFA and Culture with DFA): Sensitivity 100% (4/4); Specificity 98.6% (363/368).

Uncategorized Lesions (N=148 for HSV-1, N=166 for HSV-2, N=119 for VZV):

HSV-1 (Comparator: ELVIS ® HSV ID and D3 Typing Test): Sensitivity 100% (22/22); Specificity 97.6% (120/123).
HSV-2 (Comparator: ELVIS® HSV ID and D3 Typing Test): Sensitivity 100% (18/18); Specificity 96.6% (143/148).
VZV (Comparator: DSFA and Culture with DFA): Sensitivity 100% (17/17); Specificity 94.1% (96/102).

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

Cutaneous Lesions - HSV-1:
Sensitivity: 100% (22/22)
Specificity: 97.8% (222/227)
Cutaneous Lesions - HSV-2:
Sensitivity: 92.3% (24/26)
Specificity: 94.4% (235/249)
Cutaneous Lesions - VZV:
Sensitivity: 100% (22/22)
Specificity: 96.5% (193/200)
Mucocutaneous Lesions - HSV-1:
Sensitivity: 100% (113/113)
Specificity: 96.4% (374/388)
Mucocutaneous Lesions - HSV-2:
Sensitivity: 99.1% (108/109)
Specificity: 97.2% (487/501)
Mucocutaneous Lesions - VZV:
Sensitivity: 100% (4/4)
Specificity: 98.6% (363/368)
Uncategorized Lesions - HSV-1:
Sensitivity: 100% (22/22)
Specificity: 97.6% (120/123)
Uncategorized Lesions - HSV-2:
Sensitivity: 100% (18/18)
Specificity: 96.6% (143/148)
Uncategorized Lesions - VZV:
Sensitivity: 100% (17/17)
Specificity: 94.1% (96/102)

Predicate Device(s)

K133448

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.3309 Herpes virus nucleic acid-based cutaneous and mucocutaneous lesion panel.

(a)
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.(b)
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 820.30 design controls 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.

Summary

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized caduceus symbol, which is a staff with two snakes entwined around it, and three human profiles facing right. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the symbol.

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

November 28, 2016

Ouidel Corporation Ronald H. Lollar Senior Director, Clinical and Regulatory Affairs 2005 East State Street, Suite 100 Athens, OH 45701

Re: K162451

Trade/Device Name: Solana® HSV 1+2/VZV Assay Regulation Number: 21 CFR 866.3309 Regulation Name: Herpes virus nucleic acid-based cutaneous and mucocutaneous lesion panel Regulatory Class: II Product Code: PGI Dated: August 31, 2016 Received: September 1, 2016

Dear Mr. Lollar:

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. Isting 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 of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the

1

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 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) K162451

Device Name Solana® HSV 1+2/VZV Assay

Indications for Use (Describe)

The Solana® HSV 1+2/VZV Assay is an in vitro diagnostic test, using isothermal amplification technology (helicasedependent amplification, HDA), for the qualitative detection and differentiation of herpes simplex virus type 1, herpes simplex virus type 2, and varicella-zoster virus DNA isolated and purified from cutaneous lesion samples obtained from symptomatic patients suspected of active herpes simplex virus 1, herpes simplex virus 2 and/or varicella-zoster infection. The Solana® HSV 1+2/VZV Assay is intended to aid in the diagnosis of herpes virus 1. herpes simplex virus 2 and varicella-zoster virus active cutaneous infections. Negative results do not preclude herpes simplex virus 1, herpes simplex virus 2 and varicella-zoster virus infections and should not be used as the sole basis for diagnosis, treatment or other management decisions. The Solana® HSV 1+2/VZV Assay is intended for use only with the Solana® instrument.

Warning: The Solana® HSV 1+2/VZV Assay 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). The Solana® HSV 1+2/VZV Assay is not intended for use in prenatal screening.

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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4

Applicant:

Quidel Corporation 12544 High Bluff Drive, Suite 200 San Diego, California 92130 Telephone: 858-552-7910 Fax: 858-646-8045

Contact Information:

Ronald H. Lollar, Senior Director Clinical and Regulatory Affairs 2005 East State Street, Suite 100 Athens, Ohio 45701 740-589-3300 - Corporate number 740-589-3373 – Desk phone 858-552-6451—Fax Ron.Lollar@quidel.com

Date of preparation of 510(k) summary:

August 31, 2016

A. 510(k) Number:

K162451

B. Purpose for Submission:

To obtain substantial equivalence for the Solana® HSV 1+2/VZV Assay when performed on the Solana® instrument

C. Measurand:

HSV-1: US7: glycoprotein I

HSV-2: noncoding region between UL47: VP13/14 and UL48: VP16

VZV: ORF6: DNA-helicase primase

D. Type of Test:

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Helicase-dependent amplification (HDA)

E. Applicant:

Quidel Corporation

F. Proprietary and Established Names:

Solana® HSV 1+2/VZV Assay

G. Regulatory Information:

| Table 1.

Regulatory Information
Product Code	Classification	Regulation Section	Panel
PGI — Herpes
Virus (VZV, HSV-
1, HSV-2) DNA
Detection Assay
for Cutaneous
and
Mucocutaneous
Lesion Samples	Class II
(Special
Controls)	21 CFR 866.3309 – Herpes virus
nucleic acid-based cutaneous
and mucocutaneous lesion panel	Microbiology
(83)

H. Intended Use:

1. Intended Use(s):

The Solana® HSV 1+2/VZV Assay is an in vitro diagnostic test, using isothermal amplification technology (helicase-dependent amplification, HDA), for the qualitative detection and differentiation of herpes simplex virus type 1, herpes simplex virus type 2, and varicella-zoster virus DNA isolated and purified from cutaneous or mucocutaneous lesion samples obtained from symptomatic patients suspected of active herpes simplex virus 1, herpes simplex virus 2 and/or varicella-zoster infection. The Solana® HSV 1+2/VZV Assay is intended to aid in the diagnosis of herpes simplex virus 1, herpes simplex virus 2 and varicella-zoster virus active cutaneous or mucocutaneous infections. Negative results do not preclude herpes simplex virus 1, herpes simplex virus 2 and varicella-zoster virus infections and should not be used as the sole basis for diagnosis, treatment or other management decisions.

6

The Solana® HSV 1+2/VZV Assay is intended for use only with the Solana® instrument.

Warning: The Solana® HSV 1 + 2/VZV Assay 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). The Solana® HSV 1 + 2/VZV Assay is not intended for use in prenatal screening.

2. Indication(s) for Use:

Same as intended Use

1. Special conditions for use statement(s):
- For in vitro diagnostic use only
- For prescription use only
1. Special instrument requirements:

Solana® instrument

l. Device Description:

The Solana® HSV 1+2/VZV Assay amplifies and detects viral DNA isolated from cutaneous or mucocutaneous lesion samples obtained from symptomatic patients suspected of active herpes simplex virus 1, herpes simplex virus 2 and/or varicella-zoster infection.

The assay consists of two (2) major steps: 1) specimen preparation, and 2) amplification and detection of target sequence specific to HSV-1, HSV-2 and/or VZV using isothermal Helicase-Dependent Amplification (HDA) in the presence of target-specific fluorescence probe.

Patient specimen is transferred to a Process Tube, subjected to heat treatment at 95°C for 5 minutes and mixed and vortexed. The processed sample is transferred to a Reaction Tube and mixed. The Reaction Tube contains lyophilized HDA reagents, dNTPs, primers and probes. Once rehydrated with the diluted sample, the Reaction Tube is placed in Solana for amplification and detection of specific target sequence. In Solana, the target sequences are amplified by HSV-1, HSV-2 and/or VZV specific primers and detected by HSV-1, HSV-2 and/or VZV specific fluorescence probes included in the Reaction Tube. A competitive process control (PRC) is included in the Process Tube to monitor sample processing, inhibitory substances in clinical samples, reagent failure or device failure. The PRC target is amplified by specific primers and detected by a PRC specific fluorescence probe.

7

The target and PRC probes are labeled with a quencher on one end and a fluorophore on the other end. In addition, the target and PRC probes carry a ribonucleic acid. Upon annealing to HSV-1, HSV-2, VZV or PRC amplicons, the fluorescence probes are cleaved by RNaseH2 and the fluorescence signal increases due to physical separation of fluorophore from quencher. Solana measures and interprets the fluorescent signal, using on-board method-specific algorithms. Solana will then report the test results to the user on its display screen, and the results can be printed via an attached printer.

Materials Provided:

Cat. #M302

48 Tests per Kit

| Table 2.

Kit Components
Component	Quantity	Storage
Process Buffer Tubes	48 tubes/kit 1.6 mL	2°C to 8°C
Reaction Tubes	48 tubes/kit	2°C to 8°C

Materials required but not provided:

. External controls for HSV-1, HSV-2, or VZV (e.g. Quidel Molecular HSV/VZ Control Set, which contains positive and negative controls, serves as an external processing and extraction control)
Sterile DNase-free filter-blocked or positive displacement micropipettor tips ●
Micropipettor ●
Stopwatch or timer ●
Scissors or a blade ●
Heat block capable of 95° C ± 2° C temperature ●
Solana workflow tray and transfer rack ●
Solana Instrument
Thermometer

Substantial Equivalence Information: J.

1. Predicate device name(s):
  Lyra® Direct HSV 1+2/VZV Assay
1. Predicate 510(k) number(s): K133448

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3. Comparison with predicate:

Table 3. Similarities
Item	Subject Device
Solana® HSV 1+2/VZV Assay	Subject Device
Lyra® Direct HSV 1 + 2/VZV
Assay K133448
Intended Use	The Solana® HSV 1+2/VZV Assay is an in vitro diagnostic test, using isothermal amplification technology (helicase-dependent amplification, HDA), for the qualitative detection and differentiation of herpes simplex virus type 1, herpes simplex virus type 2, and varicella-zoster virus DNA isolated and purified from cutaneous or mucocutaneous lesion samples obtained from symptomatic patients suspected of active herpes simplex virus 1, herpes simplex virus 2 and/or varicella-zoster infection. The Solana® HSV 1+2/VZV Assay is intended to aid in the diagnosis of herpes simplex virus 1, herpes simplex virus 2 and varicella-zoster virus active cutaneous or mucocutaneous infections. Negative results do not preclude herpes simplex virus 1, herpes simplex virus 2 and varicella-zoster virus infections and should not be used as the sole basis for diagnosis, treatment or other management decisions. The Solana® HSV 1+2/VZV Assay is intended for use only with the Solana® instrument.

Warning: The Solana® HSV 1 + 2/VZV Assay is not intended for use with cerebrospinal fluid or to aid in the diagnosis of HSV or VZV infections of | The Lyra® Direct HSV 1 + 2/VZV Assay is an in vitro multiplex Real-Time PCR test for qualitative detection and differentiation of herpes simplex virus type 1, herpes simplex virus type 2, and varicella-zoster virus DNA isolated and purified from cutaneous or mucocutaneous lesion samples obtained from symptomatic patients suspected of active herpes simplex virus 1, herpes simplex virus 2 and/or varicella-zoster infection. The Lyra® Direct HSV 1 + 2/VZV Assay is intended to aid in the diagnosis of herpes simplex virus 1, herpes simplex virus 2 and varicella-zoster virus active cutaneous or mucocutaneous infections. Negative results do not preclude herpes simplex virus 1, herpes simplex virus 2 and varicella-zoster virus infections and should not be used as the sole basis for diagnosis, treatment or other management decisions. The Lyra® Direct HSV 1 + 2/VZV Assay is not intended for use with cerebral spinal fluid. The |
| Table 3. Similarities | | |
| Item | Subject Device
Solana® HSV 1+2/VZV Assay | Subject Device
Lyra® Direct HSV 1 + 2/VZV
Assay K133448 |
| | the central nervous system. The
Solana® HSV 1 + 2/VZV Assay is not
intended for use in prenatal
screening. | Lyra® Direct HSV 1 + 2/VZV
Assay is not intended for use
in prenatal screening. The
device is not intended for
point-of-care use. |
| Detection
Techniques | Multiplex assay using different
reporter dyes for each target | Multiplex assay using
different reporter dyes for
each target |
| Identification of
HSV-1, HSV-2, and
VZV | Yes | Yes |
| Assay Results | Qualitative | Qualitative |
| Sample Types | cutaneous or mucocutaneous lesion
swab specimens obtained from
symptomatic patients | cutaneous or mucocutaneous
lesion swab specimens
obtained from symptomatic
patients |
| Extraction Methods | Not required | Not required |

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	Table 4. Differences
Item	Subject Device
Solana® HSV 1+2/VZV Assay	Subject Device
Lyra® Direct HSV 1 + 2/VZV
Assay K133448
Viral Target	HSV-1: US7: glycoprotein I	HSV-1: glycoprotein G
	HSV-2: noncoding region between
UL47: VP13/14 and UL48: VP16	HSV-2: glycoprotein G
	VZV: ORF6: DNA-helicase primase	VZV: ORF6: DNA-helicase
primase

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	Table 4. Differences
Item	Subject Device
Solana® HSV 1+2/VZV Assay	Subject Device
Lyra® Direct HSV 1 + 2/VZV
Assay K133448
Assay Methodology	Helicase-Dependent Amplification
(HDA) detecting the presence or
absence of viral DNA in clinical
specimens	PCR-based system for
detecting the presence or
absence of viral DNA in
clinical specimens
Amplification and
Detection
instruments	Solana® instrument	Life Technologies
QuantStudio™ Dx, the
Applied Biosystems® 7500
Fast Dx, or the Cepheid
SmartCycler® II System

K. Standard/Guidance Document Referenced (if applicable):

Not applicable

L. Test Principle:

Patient specimen is transferred to a Process Tube, subjected to heat treatment at 95°C for 5 minutes and mixed and vortexed. The processed sample is transferred to a Reaction Tube and mixed. The Reaction Tube contains lyophilized HDA reagents, dNTPs, primers and probes. Once rehydrated with the diluted sample, the Reaction Tube is placed in Solana for amplification and detection of specific target sequence. In Solana, the target sequences are amplified by HSV-1, HSV-2 and/or VZV specific primers and detected by HSV-1, HSV-2 and/or VZV specific fluorescence probes included in the Reaction Tube. A competitive process control (PRC) is included in the Process Tube to monitor sample processing, inhibitory substances in clinical samples, reagent failure or device failure. The PRC target is amplified by specific primers and detected by a PRC specific fluorescence probe.

The target and PRC probes are labeled with a quencher on one end and a fluorophore on the other end. In addition, the target and PRC probes carry a ribonucleic acid. Upon annealing to HSV-1, HSV-2, VZV or PRC amplicons, the fluorescence probes are cleaved by RNaseH2 and the fluorescence signal increases due to physical separation of fluorophore from quencher. Solana measures and interprets the fluorescent signal, using on-board

11

method-specific algorithms. Solana will then report the test results to the user on its display screen, and the results can be printed via an attached printer.

M. Performance Characteristics:

1. Analytical performance:

a. Precision/Reproducibility:

Reproducibility

The reproducibility of the Solana HSV 1+2/VZV Assay was evaluated at three laboratory sites. A reproducibility panel containing 30 contrived samples, manufactured as high negative samples (n=3; 1/18x or 1/27x LOD (C₂o – ၆၈ concentration)) for HSV-1 MacIntyre, HSV-2 G, and VZV Ellen, low positive samples (n=3; near the assay limit of detection) for HSV-1, HSV-2 and VZV, moderate positive samples (n=3; 3x LOD) for HSV-1, HSV-2 and VZV and negative samples (n=3) was used for the study. The samples were randomized and blind-coded within each panel, and the operator tested one (1) panel, together with three (3) positive and three (3) negative external controls, in three runs. The panels were run by two operators at each testing site for five (5) non-consecutive days.

Table 5. Reproducibility Summary
	SITE						Overall Percent Agreement
HSV-1 MacIntyre	Site #1		Site #2		Site #3				95% Confidence Interval
	Rate of Detection	% Agreement	Rate of Detection	% Agreement	Rate of Detection	% Agreement	Rate of Detection	% Agreement
High Negative
(3.50× 101 TCID50/mL)	11/30	36.7	5/30	16.7	14/30	46.7	30/90	33.3	24.5 to 43.0
Low Positive
(6.30 × 102 TCID50/mL)	30/30	100	30/30	100	30/30	100	90/90	100	95.9 to 100
Moderate Positive
(1.89 × 103 TCID50/mL)	30/30	100	30/30	100	30/30	100	90/90	100	95.9 to 100
Negative	0/30	100	0/30	100	0/30	100	0/90	100	95.9 to 100
Positive Control	30/30	100	30/30	100	30/30	100	90/90	100	95.9 to 100
Negative Control	0/30	100	0/30	100	0/30	100	0/90	100	95.9 to 100

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| SITE | | | Overall Percent
Agreement | | | | | | |
|----------------------------------------------|----------------------|----------------|------------------------------|----------------|----------------------|----------------|------------------------------|----------------|-------------------------------|
| HSV-2 G | Site #1 | | Site #2 | | Site #3 | | Overall Percent
Agreement | | 95%
Confidence
Interval |
| | Rate of
Detection | %
Agreement | Rate of
Detection | %
Agreement | Rate of
Detection | %
Agreement | Rate of
Detection | %
Agreement | |
| High Negative
(3.71 × 102 TCID50/mL) | 12/30 | 40 | 8/30 | 26.7 | 7/30 | 23.3 | 27/90 | 30.0 | 21.5 to 40. |
| Low Positive
(6.67 × 103 TCID50/mL) | 30/30 | 100 | 30/30 | 100 | 30/30 | 100 | 90/90 | 100 | 95.9 to 100 |
| Moderate Positive
(2.00 × 104 TCID50/mL) | 30/30 | 100 | 30/30 | 100 | 30/30 | 100 | 90/90 | 100 | 95.9 to 10 |
| Negative | 0/30 | 100 | 0/30 | 100 | 0/30 | 100 | 0/90 | 100 | 95.9 to 100 |
| Positive Control | 30/30 | 100 | 30/30 | 100 | 30/30 | 100 | 90/90 | 100 | 95.9 to 100 |
| Negative Control | 0/30 | 100 | 0/30 | 100 | 0/30 | 100 | 0/90 | 100 | 95.9 to 100 |
| SITE | | | Overall Percent
Agreement | | | | | | |
| VZV Ellen | Site #1 | | Site #2 | | Site #3 | | Overall Percent
Agreement | | 95%
Confidence
Interval |
| | Rate of
Detection | %
Agreement | Rate of
Detection | %
Agreement | Rate of
Detection | %
Agreement | Rate of
Detection | %
Agreement | |
| High Negative
(5.50 × 10-3 TCID50/mL) | 11/30 | 36.7 | 2/30 | 6.7 | 8/30 | 26.7 | 21/90 | 23.3 | 15.8 to 33. |
| Low Positive
(1.49 × 10-1 TCID50/mL) | 30/30 | 100 | 30/30 | 100 | 30/30 | 100 | 90/90 | 100 | 95.9 to 100 |
| Moderate Positive
(4.46 × 10-1 TCID50/mL) | 30/30 | 100 | 30/30 | 100 | 30/30 | 100 | 90/90 | 100 | 95.9 to 100 |
| Negative | 0/30 | 100 | 0/30 | 100 | 0/30 | 100 | 0/90 | 100 | 95.9 to 100 |
| Positive Control | 30/30 | 100 | 30/30 | 100 | 30/30 | 100 | 90/90 | 100 | 95.9to 100 |

The results suggest that there are no significant differences between different users using different instruments at different sites on different days.

b. Linearity/assay reportable range:
Not applicable – This assay is qualitative.
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
Traceability:

Not applicable. This assay is qualitative.

Specimen Stability:

13

A study was performed to demonstrate specimen stability in various transport media types: M4, M4RT, M5, M6, and UTM when stored at room temperature (30 °C) for up to 48 hours, ≤ -20 °C, or 2 to 8 °C for up to 7 days.

HSV-1, HSV-2, and VZV virus stocks (HSV-1 McIntyre, HSV-2 G, and VZV Ellen) were diluted to their respective 2x LOD concentrations in each in of five (5) transport medium pooled negative matrix (Remel M4, Remel M6, Remel M6, Remel M4RT or UTM transport medium).

The transport media systems containing the contrived samples were stored T at three different conditions: Condition 1 = room temperature (30 ± 2 °C) for 50 hours, Condition 2 = 2 to 8 °C for 8 days, and Condition 3 = -20 °C for 8 days.

For Condition 1, the specimens were stored at room temperature (30 ± 2 °C) and tested at 0, 24, 48, and 50 hours post-storage. For Condition 2, the specimens were stored at 2-8 °C and tested at Day 0, Day 1, Day 3, Day 4, Day 7, and Day 8 poststorage. For Condition 3, the specimens were stored at -20 °C and tested at Day 0, Day 1, Day 3, Day 4, Day 7, and Day 8 post-storage.

Based on this study, HSV-1, HSV-2, and VZV samples at 2x LOD were stable when stored in M4, M4-RT, M5, M6, and UTM negative matrix for up to 50 hours at 30 °C ± 2 °C. HSV-1, HSV-2, and VZV samples were stable when stored in M4, M4-RT, M5, M6, and UTM negative matrix for up to 8 days at 2° to 8 °C. HSV-1, HSV-2, and VZV samples were stable when stored in M4, M4-RT, M5, M6, and UTM negative matrix for up to 8 days at 2° to 8 °C.

Controls:

. External controls for HSV-1, HSV-2, or VZV (e.g. Quidel Molecular HSV/VZV Control Set, which contains positive and negative controls, serves as an external processing and extraction control) were run on the Solana HSV 1+2/VZV Assay each day of testing.

The assay controls are described as follows:

a. The process control is used to monitor sample processing, to detect HDA inhibitory specimens and to confirm the integrity of assay reagents and Solana instrument functionality. The process control is included in the Process Buffer tube.

14

The external positive control may be treated as a patient specimen. The b. control should be sampled and tested as if it were a specimen and processed as described in the Assay Procedure. The external positive control is intended to monitor substantial reagent and instrument failure.
c. The external negative control may be treated as a patient specimen. The control should be sampled and tested as if it were a specimen and processed as described in the Assay Procedure. The external negative control is used to detect reagent or environmental contamination (or carry-over) by HSV 1+2/VZV DNA or amplicon.

d. Detection limit:

The analytical sensitivity (limit of detection or LOD) of the Solana HSV 1+2/VZV Assay was determined using quantified (TCID50/mL) cultures of two (2) HSV-1 strains, two (2) HSV-2 strains, and two (2) VZV strains, serially diluted in negative matrix. Each dilution was run as 20 replicates in the Solana HSV 1+2/VZV assay. Analytical sensitivity (LOD) is defined as the lowest concentration at which at least 95% of all replicates tested positive. The demonstrated LOD for each strain tested is shown below:

Table 6.	LOD Values
Virus	TCID50/mL
HSV-1 MacIntyre	6.30 × 102
HSV-1 316	5.47 × 104
HSV-2 G	6.67 × 103
HSV-2 COMP	1.62 × 105
VZV Ellen	1.49 × 10-1
VZV 9939	1.65 × 102

e. Analytical specificity:

Cross Reactivity:

A study was performed to evaluate the performance of the Solana HSV 1+2/VZV Assay in the presence of sixty-four (64) organisms that might be found in lesion specimens. Each potentially cross-reactive microorganism was tested at clinically relevant levels of viruses and bacteria: ≥10° CFU/mL; viruses: ≥ 10° copies (cp), viral particles (vp) or TCID50/mL.

15

| Table 7.

Potentially Cross-reactive Organisms
Organism	Test
Concentration	Organism	Test
Concentration
Acholeplasma laidlawi	7.10E+06 CFU/mL	Klebsiella pneumoniae	1.61E+06 CFU/mL
Acinetobacter calcoaceticus	9.27E+06 CFU/mL	Lactobacillus acidophilus	2.49E+06 CFU/mL
Adenovirus 7	1.58E+05 TCID50/mL	Legionella pneumophila	1.76E+06 CFU/mL
Bacteroides fragilis	1.19E+06 CFU/mL	Measles virus	1.95E+05 TCID50/mL
Bordetella bronchiseptica	1.97E+06 CFU/mL	Mobiluncus mulieris	2.54E+06 CFU/mL
Bordetella pertussis	7.21E+06 CFU/mL	Moraxella cartarrhalis	1.26E+06 CFU/mL
Candida albicans	2.00E+06 CFU/mL	Mumps virus	5.89E+05 TCID50/mL
Candida glabrata	3.93E+06 CFU/mL	Mycoplasma hominis	1.30E+06 CFU/mL
Chlamydia trachomatis	3.00E+06 CFU/mL	Mycoplasma hyorhinis	6.60E+06 CFU/mL
Chlamydophila pneumoniae	1.25E+06 IFU/mL	Mycoplasma orale	3.08E+06 CFU/mL
Clostridium perfringens	1.06E+06 CFU/mL	Mycoplasma pneumoniae	3.16E+06 CFU/mL
Coronavirus OC43	8.51E+05 TCID50/mL	Mycoplasma salivarium	1.67E+06 CFU/mL
Corynebacterium diphtheriae	1.51E+06 CFU/mL	Neisseria gonorrhoeae	1.23E+06 CFU/mL
Coxsackievirus B4	3.16E+05 TCID50/mL	Parainfluenza Type 1	3.97E+05 TCID50/mL
Cytomegalovirus Towne VR-977	2.14E+05 TCID50/mL	Parainfluenza Type 2	3.15E+05 TCID50/mL
Echovirus 11	2.14E+05 TCID50/mL	Parainfluenza Type 3	2.56E+05 TCID50/mL
Enterococcus faecalis	3.45E+06 CFU/mL	Parainfluenza Type 4	1.37E+05 TCID50/mL
Enterovirus 70	1.78E+05 TCID50/mL	Proteus mirabilis	1.19E+06 CFU/mL
Epstein Barr Virus	1.34E+05 vp/mL	Pseudomonas aeruginosa	1.32E+06 CFU/mL
Table 7. Potentially Cross-reactive Organisms
Organism	Test Concentration	Organism	Test Concentration
Escherichia coli	8.42E+06 CFU/mL	RSV A Long	1.95E+05 TCID50/mL
Gardnerella vaginalis	1.20E+06 CFU/mL	RSV B Washington	3.43E+05 TCID50/mL
Haemophilis influenza type A	5.33E+06 CFU/mL	Rubella Virus	2.09E+05 TCID50/mL
HBV synthetic DNA	6.80E+05 cp/mL	Salmonella enteriditis	5.40E+06 CFU/mL
HCV synthetic RNA	1.96E+05 cp/mL	Salmonella typhimurium	1.01E+06 CFU/mL
HHV-6	3.30E+05 TCID50/mL	Staphylococcus aureus	1.02E+06 CFU/mL
HHV-7	1.15E+05 TCID50/mL	Staphylococcus saprophyticus	2.00E+06 CFU/mL
HHV-8	1.26E+05 TCID50/mL	Streptococcus agalactiae	2.20E+06 CFU/mL
HIV purified RNA	1.60E+05 cp/mL	Streptococcus pneumoniae	2.18E+06 CFU/mL
hMPV A1	3.66E+05 TCID50/mL	Streptococcus pyogenes	1.29E+06 CFU/mL
HPV	4.30E+05 cp/uL	Toxoplasma gondii	1.06E+06 tachyzoites/mL
Influenza A/Mexico/4108/2009	2.88E+05 vp/mL	Trichomonas vaginalis	1.00E+06 trophozoites/mL
Influenza B Hong Kong VR-791	1.91E+05 TCID50/mL	Ureaplasma uralyticum	1.23E+06 CFU/mL

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No cross-reactivity was observed with the sixty-four (64) microorganisms tested with the Solana HSV 1+2/VZV Assay. Interference:

The performance of Solana HSV 1+2/VZV Assay was evaluated with potentially interfering substances that may be present in lesion specimens. A panel composed of twenty-six (26) substances was tested in the absence or presence of HSV-1, HSV-2, or VZV (Maclntyre, G, Ellen strains, respectively) at 2X LOD in the Solana HSV 1+2/VZV Assay. There was no evidence of interference caused by the substances tested at the concentrations shown below.

17

| Table 8.

Potential Interfering Substances and Concentrations Tested
Substance	Test
Concentration	Substance	Test
Concentration
Abreva	7%	Female Urine	7%
Acetamidophenol	10 mg/mL	KY Jelly	7%
Acyclovir	7 mg/mL	Lanacane	3.50%
Albumin	3.3 mg/mL	Leukocytes	2.5x105 cells/mL
Blood/EDTA	0.63%	Listerine	7%
Carmex	7%	Male Urine	7%
Casein	7 mg/mL	Miconazole 1	7%
Chlorpheniramine	5 mg/mL	Miconazole 3	7%
Colgate	7%	Mucin	60 µg/mL
Cornstarch	2.5 mg/mL	Preparation H	7%
Dextromethorphan	5 mg/mL	Releev	7%
Douche	7%	Seminal Fluid	2%
Feces	0.22%	Tioconazole 1	7%

Analytical Reactivity (Inclusivity):

The inclusivity of the Solana HSV 1+2/VZV Assay was further evaluated by functional testing of viral strains in addition to those strains used in the LOD study. The clinical panel consisted of two (2) strains of HSV-1, three (3) strains of HSV-2, and five (5) strains of VZV at concentrations near the level of detection (LOD) of the assay.

Table 9. Inclusivity Strains
Strain	TCID50/mL	Inclusive (Yes or No)
HSV-1 Isolate #1	1.26 × $10^3$	Yes
HSV-1 Isolate #3	1.26 × $10^3$	Yes
HSV-2 Strain MS	1.33 × $10^4$	Yes
HSV-2 Isolate #25	1.33 × $10^4$	Yes

18

Table 9. Inclusivity Strains
Strain	TCID50/mL	Inclusive (Yes or No)
HSV-2 Isolate #32	1.33 × 104	Yes
VZV Strain 82	8.05 × 100	Yes
VZV Strain 130	2.41 × 101	Yes
VZV Strain 275	8.05 × 100	Yes
VZV Strain B	2.41 × 101	Yes
VZV Strain D	8.05 × 100	Yes

f. Assay cut-off:
Not applicable.

2. Comparison studies:

a. Method comparison with predicate device:
Not applicable
b. Matrix comparison:
Not applicable
1. Clinical studies:
- Clinical Sensitivity: a.

Performance characteristics of the Solana " HSV 1+2/VZV Assay were established during a prospective study between February and April 2016. One thousand sixtytwo (1062) fresh lesion specimens, collected for herpes simplex/varicella-zoster identification, have been included in this study at three sites across the United States. A single specimen was collected per patient. The specimens were processed and tested with Solana " HSV 1+2/VZV Assay on the Solana "instrument at the sites. Testing of the comparator methods (culture with DFA) were performed at one central location.

19

Combined Data

Cutaneous Lesions

Two-hundred seventy-five (275) active cutaneous lesion specimens were cultured for HSV-1, using the FDA-cleared ELVIS cell culture system and were also tested with the subject device for HSV-1 viral DNA. Samples which gave HSV-2 positive results from the ELVIS method were excluded from the HSV-1 performance calculation because of the inability of the ELVIS method to distinguish an HSV-1 positive sample when HSV-2 was detected first (n=26). The table below details the HSV-1 results for the remaining two-hundred forty-nine (249) specimens.

Table 10. HSV-1 Results
Comparator: ELVIS® HSV ID and D3 Typing Test
Solana HSV 1 + 2/VZV Assay	Positive	Negative	Total
Positive	22	5*	27
Negative	0	222	222
Total	22	227	249
95% CI
Sensitivity	22/22	100%	85.1% to 100%
Specificity	222/227	97.8%	95.0% to 99.1%

Three (3) of the five (5) positives was positive by an additional RT-PCR assay.

Two-hundred seventy-five (275) active cutaneous lesion specimens were cultured for HSV-2, using the FDA-cleared ELVIS cell culture system and were also tested with the subject device for HSV-2 viral DNA. The table below details the HSV-2 results for the two-hundred seventy-five (275).

	Table 11.	HSV-2 Results
Comparator: ELVIS® HSV ID and D³ Typing Test
Solana HSV 1 + 2/VZV
Assay	Positive	Negative	Total
Positive	24	14*	28
Negative	2**	235	237
Total	26	249	275
95% CI
Sensitivity	24/26	92.3%	75.9% to 97.9%
Specificity	235/249	94.4%	90.8% to 96.6%

Thirteen (13) of the fourteen (14) positives were positive by an additional RT-PCR assay.

** Two (2) of the two (2) negatives were positive by an additional RT-PCR assay.

20

Two-hundred and seventy-five (275) active cutaneous lesion specimens were cultured for VZV using the H&V mixed cells with DFA cell culture systems and were also tested with the subject device for VZV viral DNA. Due the presence of either HSV-1 or HSV-2, fifty-one (51) specimens have been excluded from analysis. Two (2) specimens were contaminated or had toxic cultures. These fifty-three (53) specimens have been excluded from analysis. The table below details the VZV results for the remaining two-hundred and twenty-two (222) specimens.

	Table 12.	VZV Results
Comparator: DSFA and Culture with DFA
Solana HSV 1 + 2/VZV Assay	Positive	Negative	Total
Positive	22	7*	29
Negative	0	193	193
Total	22	200	222
			95% CI
Sensitivity	22/22	100%	85.1% to 100%
Specificity	193/200	96.5%	93.0% to 98.3%

Six (6) of the seven (7) positives were positive by an additional RT-PCR assay.

Mucocutaneous Lesions

Six-hundred twenty-one (621) active mucocutaneous lesion specimens were cultured for HSV-1, using the FDA-cleared ELVIS cell culture system and were also tested with the subject device for HSV-1 viral DNA. Seven (7) specimens were contaminated in the ELVIS cell culture. Two (2) specimens were invalid in Solana HSV 1 + 2/VZV Assay. Two (2) specimens were contaminated and invalid. Samples which gave HSV-2 positive results from the ELVIS method were excluded from the HSV-1 performance calculation because of the inability of the ELVIS method to distinguish an HSV-1 positive sample when HSV-2 was detected first (n=109). Thus, one hundred and twenty (120) specimens have been excluded from further analysis. The table below details the HSV-1 results for the remaining five-hundred and one (501) specimens.

21

	Table 13.	HSV-1 Results
Comparator: ELVIS® HSV ID and D3 Typing Test
Solana HSV 1 + 2/VZV Assay	Positive	Negative	Total
Positive	113	14*	127
Negative	0	374	374
Total	113	388	501
95% CI
Sensitivity	113/113	100%	96.7% to 100%
Specificity	374/388	96.4%	94.0% to 97.8%

Six (6) of the fourteen (14) positives were positive by an additional RT-PCR assay.

Six-hundred twenty-one (621) active mucocutaneous lesion specimens were cultured for HSV-2, using the FDA-cleared ELVIS cell culture system and were also tested with the subject device for HSV-2 viral DNA. Seven (7) specimens were contaminated in the ELVIS cell culture. Two (2) specimens were invalid in Solana HSV 1 + 2/VZV Assay. Two (2) specimens were contaminated and invalid. These eleven (11) specimens have been excluded from further analysis. The table below details the HSV-2 results for the remaining six-hundred ten (610) specimens.

	Table 14.	HSV-2 Results
	Comparator: ELVIS® HSV ID and D³ Typing Test
Solana HSV 1 + 2/VZV Assay	Positive	Negative	Total
Positive	108	14*	122
Negative	1**	487	488
Total	109	501	610
95% CI
Sensitivity	108/109	99.1%	95.0% to 99.8%
Specificity	487/501	97.2%	95.4% to 98.3%

Eleven (11) of the fourteen (14) positives were positive by an additional RT-PCR assay.

** One (1) of one (1) negative was positive by an additional RT-PCR assay.

Six-hundred twenty one (621) active mucocutaneous lesion specimens were cultured for VZV using the H&V mixed cells with DFA cell culture systems and were also tested with the subject device for VZV viral DNA. Due the presence of either HSV-1 or HSV-2, two hundred thirty-six (236) specimens have been excluded from analysis. Nine (9) specimens were contaminated in culture, and four (4) specimens were invalid in Solana HSV 1 + 2/VZV Assay. These two hundred forty-nine (249) specimens have been excluded from analysis. The

22

table below details the VZV results for the remaining three hundred seventy-two (372) specimens.

Table 15. VZV Results
Comparator: DSFA and Culture with DFA
Solana HSV 1 + 2/VZV Assay	Positive	Negative	Total
Positive	4	5*	9
Negative	0	363	363
Total	4	368	372
95% CI
Sensitivity	4/4	100%	51.0% to 100%
Specificity	363/368	98.6%	96.9% to 99.4%

One (1) of the five (5) positives was positive by an additional RT-PCR assay.

Note: The data presented for the detection of VZV is consistent with limited presence of VZV in mucocutaneous lesions. The use of mucocutaneous lesions has no discernible impact on the performance characteristics of Solana HSV 1 + 2/VZV Assay.

Uncategorized Lesions

One hundred sixty-six (166) active lesion specimens (not categorized as cutaneous or mucocutaneous) were cultured for HSV-1, using the FDA-cleared ELVIS cell culture system and were also tested with the subject device for HSV-1 viral DNA. Samples which gave HSV-2 positive results from the ELVIS method were excluded from the HSV-1 performance calculation because of the inability of the ELVIS method to distinguish an HSV-1 positive sample when HSV-2 was detected first (n=18). The table below details the HSV-1 results for the remaining one hundred and forty-eight (148) specimens.

	Table 16.	HSV-1 Results
	Comparator: ELVIS ® HSV ID and D3 Typing Test
Solana HSV 1 + 2/VZV
Assay	Positive	Negative	Total
Positive	25	3*	28
Negative	0	120	120
Total	25	123	148
			95% CI
Sensitivity	22/22	100%	86.7% to 100%
Specificity	120/123	97.6%	93.1% to 99.2%

Three (3) of the three (3) positives was positive by an additional RT-PCR assay.

23

One hundred sixty-six (166) active lesion specimens (not categorized as cutaneous or mucocutaneous) were cultured for HSV-2, using the FDA-cleared ELVIS cell culture system and were also tested with the subject device for HSV-2 viral DNA. The table below details the HSV-2 results for the one hundred sixtysix (166).

	Table 17.	HSV-2 Results
Comparator: ELVIS® HSV ID and D3 Typing Test
Solana HSV 1 + 2/VZV
Assay	Positive	Negative	Total
Positive	18	5*	23
Negative	0	143	143
Total	18	148	166
95% CI
Sensitivity	18/18	100%	82.4% to 100%
Specificity	143/148	96.6%	92.3% to 98.6%

Five (5) of the five (5) positives were positive by an additional RT-PCR assay.

One hundred sixty-six (166) active lesion specimens (not categorized as cutaneous or mucocutaneous) were cultured for VZV using the H&V mixed cells with DFA cell culture systems and were also tested with the subject device for VZV viral DNA. Due the presence of either HSV-1 or HSV-2, forty-six (46) specimens have been excluded from analysis. One (1) specimen was contaminated in culture. These forty-seven (47) specimens have been excluded from analysis. The table below details the VZV results for the remaining one hundred nineteen (119) specimens.

	Table 18.	VZV Results
Comparator: DSFA and Culture with DFA
Solana HSV 1 + 2/VZV Assay	Positive	Negative	Total
Positive	17	6*	23
Negative	0	96	96
Total	17	102	119
95% CI
Sensitivity	17/17	100%	81.6% to 100%
Specificity	96/102	94.1%	87.8% to 97.3%

Five (5) of the six (6) positives were positive by an additional RT-PCR assay.

24

b. Clinical specificity:
See Section 3a.
Other clinical supportive data (when a. and b. are not applicable): ﻥ
Not applicable
1. Clinical cut-off:
  Not applicable

5. Expected values:

The expected values of the Solana HSV 1+2/VZV Assay were established during a prospective study conducted between February and May 2016. One thousand sixty-two (1062) specimens have been included in this study at three (3) sites across the United States. A single specimen was collected per patient. The specimens were processed and tested with Solana HSV 1+2/VZV Assay on the Solana instrument at the sites.

The expected value of HSV-1, HSV-2, and VZV with the Solana HSV 1+2/VZV Assay has been calculated for the combined sites based on the category of specimen (cutaneous, mucocutaneous, or uncategorized lesion source) and the age of the patient.

Table 19. Combined Study – Expected Values (Cutaneous) (N=273)
	HSV-1			HSV-2			VZV
Age	Total #	Total Positive	Prevalence	Total #	Total Positive	Prevalence	Total #	Total Positive	Prevalence
≤ 5 years	9	1	11.1%	9	0	N/A	9	4	44.4%
6 to 21 years	43	10	23.3%	43	4	9.3%	43	0	N/A
22 to 59 years	180	16	8.9%	180	26	14.4%	180	19	10.6%
≥ 60 years	43	0	N/A	43	8	18.6%	43	6	14.0%

Table 20.	Expected Values (Cutaneous) (N=273)
	HSV- 1			HSV-2			VZV
	Total #	Total
Positive	Prevalence	Total #	Total
Positive	Prevalence	Total #	Total
Positive	Prevalence
Genital - penis	103	10	9.7%	103	18	17.5%	103	1	1.0%
skin lesion	170	17	10.0%	170	19	11.2%	170	28	16.5%

25

Table 21. Combined Study – Expected Values (Mucocutaneous) (N=617)*
	HSV-1			HSV-2			VZV
Age	Total #	Total Positive	Prevalence	Total #	Total Positive	Prevalence	Total #	Total Positive	Prevalence
≤ 5 years	21	4	19.0%	21	0	N/A	21	1	4.8%
6 to 21 years	158	41	25.9%	158	29	18.4%	158	0	N/A
22 to 59 years	385	74	19.2%	385	89	23.1%	385	8	2.1%
≥ 60 years	53	11	20.8%	53	5	9.4%	53	1	1.9%

Four (4) specimens were invalid and removed from analysis

Table 22. Expected Values (Mucocutaneous) (N=619)*
	HSV-1			HSV-2			VZV
	Total

| Total

Positive | Prevalence | Total # | Total
Positive | Prevalence | Total # | Total
Positive | Prevalence |
| Anorectal | 26 | 7 | 26.9% | 26 | 7 | 26.9% | 26 | 1 | 3.8% |
| genital – vaginal/cervical | 449 | 80 | 17.8% | 449 | 112 | 24.9% | 449 | 5 | 1.1% |
| Nares | 23 | 5 | 21.7% | 23 | 1 | 4.3% | 23 | 3 | 13.0% |
| Ocular | 7 | 2 | 28.6% | 7 | 0 | N/A | 7 | 1 | 14.3% |
| Oral lesion | 112 | 36 | 32.1% | 112 | 3 | 2.7% | 112 | 0 | N/A |

Four (4) specimens were invalid and removed from analysis

Table 23. Combined Study – Expected Values (Uncategorized Lesion Source) (N=166)
	HSV-1			HSV-2			VZV
Age	Total #	Total Positive	Prevalence	Total #	Total Positive	Prevalence	Total #	Total Positive	Prevalence
≤ 5 years	11	1	9.1%	11	0	N/A	11	0	N/A
6 to 21 years	28	10	35.7%	28	0	N/A	28	2	7.1%
22 to 59 years	89	15	16.9%	89	18	20.2%	89	14	15.7%
≥ 60 years	38	2	5.3%	38	5	13.2%	38	8	21.1%

N. Other Supportive Instrument Performance Characteristics Data Not Covered In The "Performance Characteristics" Section above:

Instrument: Solana™ Instrument

O. System Descriptions:

Modes of Operation:

26

The Solana instrument heats each reaction tube to 64°C. If present, the target sequence is amplified by HSV-1, HSV-2 and/or VZV specific primers and detected by HSV-1, HSV-2 and/or VZV specific fluorescence probes included in the Reaction Tube. Each probe has a florescent dye of specific wavelength. The target probes are labeled with a quencher on one end and a fluorophore on the other end. In addition, the target probes carry a ribonucleic acid. Upon annealing to the respective amplicons, the fluorescence probes are cleaved by RNaseH2 and the fluorescence signal increases due to physical separation of fluorophore from quencher. The Solana instrument measures and interprets the fluorescent signal, using on-board method-specific algorithms. Solana instrument will then report the test results to the user on its display screen, and it can print out the results via a printer.

2. Software:

FDA has reviewed applicant's Hazard Analysis and software development processes for this line of product types:

Yes No

P. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10, 21 CFR 801.109, and the special controls.