For In Vitro Diagnostic Use Only. The SeraQuest HSV Type 1 Specific IgG assay is an enzyme-linked immunosorbent assay (ELISA) intended for the qualitative detection of human IgG antibodies to type 1 herpes simplex virus (HSV) in human serum. The test is indicated for sexually active individuals and expectant mothers as an aid in the presumptive diagnosis of HSV-1 infection. The predictive value of a positive result depends on the prevalence of HSV-1 infection in the population and the pre-test likelihood of HSV-1 infection.

The test is not FDA cleared for screening blood or plasma donors. The performance of this assay has not been established for immunocompromised patients, pediatric patients or matrices other than human serum.

Device Description

The SeraQuest® HSV Type 1 Specific IgG test is a solid-phase enzyme-linked immunoassay (ELISA), which is performed in microwells, at room temperature, and in three thirty minute incubations. The test detects IqG antibodies which are directed against HSV 1 type-specific antigens in human serum. The Calibrator in the SeraQuest® HSV Type 1 Specific IgG test set has been assigned Index values based on an in-house standard. Test results are reported as Index values.

AI/ML Overview

The SeraQuest HSV Type 1 Specific IgG assay is an enzyme-linked immunosorbent assay (ELISA) intended for the qualitative detection of human IgG antibodies to type 1 herpes simplex virus (HSV) in human serum.

Here's an analysis of the acceptance criteria and the studies performed:

Table of Acceptance Criteria and Reported Device Performance

Acceptance Criteria / Performance Metric	SeraQuest HSV Type 1 Specific IgG Performance for Sexually Active Adults	SeraQuest HSV Type 1 Specific IgG Performance for Expectant Mothers	SeraQuest HSV Type 1 Specific IgG Performance with CDC Panel
Sensitivity	92.3% (95% CI: 85.0% to 96.2%)	93.3% (95% CI: 87.3% to 96.6%)	91.3% (42/46 positive, 2 equivocal, 2 negative)
Specificity	91.7% (95% CI: 83.2% to 96.2%)	89.4% (95% CI: 87.1% to 93.7%)	98.1% (53/54 negative, 1 equivocal)

Note: The document does not explicitly state pre-defined acceptance criteria (e.g., "Sensitivity must be >= X%"). Instead, it presents the results of a comparison study against a predicate device and a CDC panel, inferring that the performance achieved is deemed acceptable for substantial equivalence.

Sample Size and Data Provenance
- Test Set (Comparative Study with Predicate Device):
  - Sexually Active Adults: 164 serum samples.
  - Expectant Mothers: 242 serum samples (198 during the first trimester, 19 during the second, 25 during the third).
  - Data Provenance: Prospectively collected, masked, and archived serum samples submitted for HSV serology to clinical laboratories in the Southeastern United States (for sexually active adults) and Northeastern and Southeastern United States (for expectant mothers). This indicates prospective data collection from the United States.
- Test Set (CDC Panel): 100 serum samples (46 HSV-1 IgG positive, 54 HSV-1 IgG negative).
  - Data Provenance: This is a characterized serum panel provided by the Centers for Disease Control and Prevention (CDC). The specific country of origin for individual samples within the CDC panel is not specified, but the panel itself is a US-based reference.
Number of Experts and Qualifications for Ground Truth
- The document does not mention the use of experts to establish ground truth for the comparative studies.
- For the comparative study, the predicate device (Focus HerpeSelect® 1 and 2 Immunoblot IgG) served as the reference standard. The ground truth was based on the results of this legally marketed predicate device.
- For the CDC panel, the CDC HSV 1 Result was used as the ground truth. This panel is composed of "well characterized serum panel" but the methods or experts used by CDC to characterize it are not detailed in this document.
Adjudication Method for the Test Set
- No adjudication method (e.g., 2+1, 3+1) is mentioned for the test set. The results are directly compared to the predicate device's findings or the CDC's characterization of the panel.
Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
- No MRMC comparative effectiveness study was done. The device is an in-vitro diagnostic (IVD) assay, not an imaging device typically evaluated with human readers. Therefore, there is no discussion of human reader improvement with or without AI assistance.
Standalone Performance
- Yes, standalone performance was done. The entire submission details the performance of the SeraQuest HSV Type 1 Specific IgG assay as a standalone device, directly comparing its results to a legally marketed predicate device (immunoblot) and a characterized CDC panel. The performance metrics (sensitivity, specificity, precision) are derived from the device's independent operation.
Type of Ground Truth Used
- Comparative Studies: The ground truth was established by another legally marketed device (predicate device): the Focus HerpeSelect® 1 and 2 Immunoblot IgG.
- CDC Panel: The ground truth was based on the characterization of the CDC serum panel, which is described as "well characterized."
Sample Size for the Training Set
- The document does not explicitly state a "training set" size. For IVD devices, the development and optimization of the assay might involve internal studies and smaller panels, but these are typically not referred to as a separate "training set" in the same way machine learning models are. The performance studies presented are generally considered validation studies on independent test sets.
How the Ground Truth for the Training Set Was Established
- As no explicit "training set" is mentioned in the context of this IVD device, the method for establishing its ground truth is not provided. The document focuses on the validation of the device against established external references.

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 circular seal with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is a stylized image of three human profiles facing right, with a symbolic representation of an eagle above them.

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

October 1, 2016

QUEST INTERNATIONAL, INC DAVID J. KIEFER, PH.D. DIRECTOR RESEARCH AND DEVELOPMENT 8127 NW 29TH STREET MIAMI FL 33122

Re: K162276

Trade/Device Name: SeraQuest HSV Type 1 Specific IgG Regulation Number: 21 CFR 866.3305 Regulation Name: Herpes simplex virus serological assays Regulatory Class: II Product Code: MXJ Dated: August 10, 2016 Received: August 12, 2016

Dear Dr. Kiefer:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug. and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21. Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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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

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Indications for Use

510(k) Number (if known) K162276

Device Name

SeraQuest® HSV Type 1 Specific IgG

Indications for Use (Describe)

Type of Use (Select one or both, as applicable)
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Prescription Use (Part 21 CFR 801 Subpart D)

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

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510(k) Summary

510(k) Number:	K162276
Purpose for Submission:	New Device
Type of Test:	Enzyme linked immunosorbent assay (ELISA)
Measurand:	IgG Antibody to Herpes Simplex Virus-1 (HSV-1)
Applicant:	Quest International, Inc.8127 NW 29th StreetDoral, FL. 33122
Contact Person:	David J. Kiefer, Ph.D.
Telephone:	305 592-6991
Telefax:	305 592-6834
Device:	SeraQuest® HSV Type 1 Specific IgG

Requlatory Information:

1. Regulation section: Herpes simplex virus serological assays 21 CFR§866.3305
1. Classification: Class II
1. Product code: MXJ (Enzyme Linked Immunosorbent Assay, Herpes Simplex Virus, HSV-1)
1. Panel: Microbiology (83)

Cataloque Number: 01-410

Intended Use:

For In Vitro Diagnostic Use Only. The SeraQuest HSV Type 1 Specific IgG assay is an enzymelinked immunosorbent assay (ELISA) intended for the qualitative detection of human IgG antibodies to type 1 herpes simplex virus (HSV) in human serum. The test is indicated for sexually active individuals and expectant mothers as an aid in the presumptive diagnosis of HSV-1 infection. The predictive value of a positive or negative result depends on the prevalence of HSV-1 infection in the population and the pre-test likelihood of HSV-1 infection.

Description:

The SeraQuest® HSV Type 1 Specific IgG test is a solid-phase enzyme-linked immunoassay (ELISA), which is performed in microwells, at room temperature, and in three thirty minute

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incubations. The test detects IqG antibodies which are directed against HSV 1 type-specific antigens in human serum. The Calibrator in the SeraQuest® HSV Type 1 Specific IgG test set has been assigned Index values based on an in-house standard. Test results are reported as Index values. The following table shows the interpretation of results:

Index	Result	Interpretation
$\leq$ 0.9	Negative	No HSV-1 IgG antibodies detected. Patient is presumed not to have had a previous HSV-1 infection.
0.9 < X < 1.0	Equivocal	Obtain an additional sample for re-testing
$\geq$ 1.0	Positive	IgG antibody to HSV-1 detected.

Notes:

1. A single positive result only indicates previous immunologic exposure: the level of antibody response may not be used to determine active infection or disease stage.
1. When equivocal results are obtained, another specimen should be obtained ten to fourteen days later, and tested in parallel with the initial specimen. If the second specimen is also equivocal, the patient is negative for primary or recent infection, and equivocal for antibody status. If the second sample is positive, the patient can be considered to have previous experience with HSV-1 infection.
1. Values obtained with different manufacturer's assay methods may not be used interchangeably. The magnitude of the reported IgG index value cannot be correlated to an endpoint titer. The magnitude of results above the cut-off is not an indicator of total antibody present.

Principle:

Diluted patient samples are incubated in antigen-coated wells. HSV Type 1 antibodies, if present in the patient sample, are immobilized in the wells by binding to the antigen. Residual sample is eliminated by washing and draining, and conjugate (enzyme-labeled antibodies to human IgG) is added and incubated. If IgG antibodies to HSV-1 are present, the conjugate will be immobilized in the wells. Residual conjugate is eliminated by washing and draining, and the substrate is added and incubated. In the presence of the conjugate, the substrate is converted to a yellow end-product which is read photometrically.

Substantial Equivalence Information:

1. Predicate device name(s): Focus HerpeSelect® 1 and 2 Immunoblot IgG
1. Predicate 510(k) number(s): K000238

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3. Comparison with predicate:

	Table 1. Comparison with the Predicate
--	----------------------------------------	--	--

Similarities
ltem	SeraQuest HSV 1 IgG	Focus HerpeSelect 1 and 2Immunoblot IgG (Predicate)
Intended Use	For In VitroDiagnostic Use Only. TheSeraQuest HSV Type 1Specific IgG assay is anenzyme-linked immunosorbentassay (ELISA) intended for thequalitative detection of humanlgG antibodies to type 1herpes simplex virus (HSV) inhuman serum. The test isindicated for sexually activeindividuals and expectantmothers as an aid in thepresumptive diagnosis ofHSV-1 infection. Thepredictive value of a positive ornegative result depends on theprevalence of HSV-1 infectionin the population andthe pre-test likelihood of HSV-1 infection.The test is not FDA clearedfor screening blood or plasmadonors. The performance ofthis assay has not beenestablished forimmunocompromisedpatients, pediatric patients ormatrices other than humanserum.	Focus Diagnostics'HerpeSelect 1 and 2Immunoblot IgG test isintended for qualitativelydetecting the presence orabsence of human IgG classantibodies to HSV-1 andHSV-2 in human sera. Thetest is indicated for testingsexually active adults orexpectant mothers for aidingin the presumptive diagnosisof HSV-1 and HSV-2infection. The predictive valueof a positive or negative resultdepends on the population'sprevalence and the pretestlikelihood of HSV-1 and HSV-2 infection.The performance of thisassay has not beenestablished for use in apediatric population, forneonatal screening, fortesting ofimmunocompromisedpatients, for use by a point ofcare facility or for use withautomated equipment.
Sample Matrix	Human Serum	Human Serum
Antigen	Recombinant HSV gG 1	Recombinant HSV gG 1
Incubation Periods	Three	Three
Incubation Temperature	Room Temperature	Room Temperature
Enzyme-Labeled Conjugate	Alkaline phosphataseconjugated goat anti-humanIgG	Alkaline phosphataseconjugated goat anti-humanIgG
Controls	Positive and negative controlsincluded with kit	Positive and negative controlsincluded with kit
Differences
Type of Assay	Enzyme linked immunoassay(ELISA)	Nitrocellulose immunoblot
Solid Phase	Polystyrene Microwells	Nitrocellulose membrane
Calibrator	Included with reagent kit	Not included
Sample Dilution	1:51	1:101
Sample Volume	100 uL	2000 uL
Sample incubation duration	30 minutes	60 minutes
Washing Steps	Two	Four
Cycles per wash step	Four	Three
Conjugate Volume	100 uL	2000 uL
Enzyme substrate	p-nitrophenyl phosphate	Bromo-chloro-indolylphosphate and nitro bluetetrazolium
Substrate Volume	100 uL	200 uL
Substrate Incubation	30 minutes	5 to 30 minutes
Stop Reagent	0.5 M Trisodium Phosphate	De-ionized Water
Stop Reagent Volume	100 uL	5X2000 uL
Drying Step	None	Air dry, 10 minutes
Readout	Spectrophotometric	Visual
Unit of Measure	Index value	Positive or negative

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Summary of Performance Testing

1. Precision Testing

Six serum specimens (2 negative, 1 equivocal and 3 positive) and the SeraQuest HSV Type 1 Specific IgG Positive and Negative Controls, were assayed in triplicate, on three separate occasions, at Quest International and at two external independent laboratories. The results are summarized below in Table 2.

Table 2.

Results of Intra-assay, Inter-assay and Inter-laboratory Precision Tests Performed at Quest International and at Two External, Independent Laboratories. The Means, Standard Deviations and Coefficients of Variation were Calculated from the SeraQuest Index Values.

Name of AnalytePanel Member	SampleN	MeanIndex	Intra-assay		Inter-assay		Inter-laboratory		Total
			SD	CV%	SD	CV%	SD	CV%	SD	CV%
SeraQuest PositiveSerum Control	27	2.0	0.12	5.8	0.13	7.2	0.17	8.5	0.14	7.2
SeraQuest NegativeSerum Control	27	0.3	0.03	10.1	0.08	23.7	0.12	39.0	0.08	24.2
Negative Sample # 1	27	0.3	0.02	7.0	0.03	10.7	0.12	43.0	0.05	20.1
Negative Sample # 2	27	0.5	0.01	2.6	0.03	5.6	0.09	17.7	0.04	8.7
Equivocal Sample	27	1.0	0.08	8.6	0.10	10.7	0.15	15.1	0.11	11.4
Positive Sample # 1	27	1.8	0.12	6.7	0.16	8.9	0.23	12.7	0.17	9.5
Positive Sample # 2	27	1.9	0.11	5.8	0.18	9.5	0.21	11.2	0.17	8.8
Positive Sample # 3	27	2.6	0.11	4.3	0.20	7.6	0.43	16.6	0.25	9.5

2. Specificity Testing

A cross-reactivity study was performed to determine if samples from various disease states and other potentially cross-reacting agents interfere with the test results. The samples were determined to be positive for IgG antibodies directed against taxonomically related viruses and other related pathogens by other legally marketed devices. Human Papilloma Virus, Chlamydia trachomitis, and Neisseria gonorrhea samples were from individual patients with confirmed sexually transmitted infections. The samples were also tested for Type 1 HSV antibody by another legally marketed device. Only those samples that tested negative for Type 1 HSV antibody by the legally marketed device were included in the study. The results of this study are shown below in Table 3.

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Table 3. Results of SeraQuest HSV Type 1 Specific IgG Tests of Samples Which Tested Positive for Antibodies Directed Against Taxonomically Related Viruses and Other Viruses and Pathogens but Negative for Type 1 HSV by Other Legally Marketed Devices.

Samples	Number of Samples	Number of Samples Testing Positive in theSeraQuest HSV 1 Type Specific IgG Test
HSV 2 IgG	10	0/10
CMV IgG	11	0/11
EBV EBNA IgG	13	0/13
EBV VCA IgG	17	0/17
VZV IgG	19	0/19
Measles IgG	20	0/20
Rubella IgG	15	0/15
Toxoplasma IgG	6	0/6
Syphilis IgG	4	0/4
Human Papilloma	2	1/2
Chlamydia trachomitis	1	0/1
Neisseria gonorrhea	3	1/3

3. Interference Testing

The effects of icterus, hemolysis, hyperglycemia, hyperlipidemia and hyperproteinemia on the test results were examined. Samples that were negative, weakly positive and moderately positive for antibodies to Type 1 HSV were tested with and without the addition of elevated levels of the following potential interfering substances: hemoglobin 18 g/dL, glucose 800 mg dL, cholesterol 2,720 mg/dL, globulin 28 g/dL, unconjugated bilirubin 20 mg/dL, conjugated bilirubin 20 mg/dL, human albumin 12 g/dL and ascorbic acid 3mg/dL.

No significant interference was observed in the presence of abnormally elevated levels of the potentially interfering substances tested, however the use of grossly hemolyzed, icteric or lipemic samples, as well as samples containing particulate matter or exhibiting obvious microbial contamination is not recommended and they should not be tested.

4. Comparison with The Predicate Device

One hundred and sixty-four serum samples, from sexually active adults that were submitted for HSV serology to a clinical laboratory in the Southeastern United States, were prospectively collected, masked, archived, and tested using the SeraQuest HSV Type 1 Specific IgG test and a commercial HSV 1 Immunoblot test. The results of this comparative test are shown in Table 4 below.

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Table 4.

Relative Sensitivity and Specificity of the SeraQuest HSV 1 Type Specific IgG Test in Comparison with an HSV 1 Immunoblot Test for 164 Sexually Active Adults Whose Sera Had Been Submitted for Herpes Simplex Virus Serology.

SeraQuest HSV 1 Type Specific IgG Result
HSV 1 Immunoblot Result	Positive	Equivocal	Negative	% Agreement*	95% C.I.*
Positive	84	1	6	Sensitivity 92.3	85.0 to 96.2
Negative	3	3	67	Specificity 91.7	83.2 to 96.2

Two hundred and forty-two serum samples, from expectant mothers, that were submitted for HSV serology to clinical laboratories in the Northeastern and Southeastern United States, were prospectively collected, masked, archived, and tested using the SeraQuest HSV Type 1 Specific IgG test and a legally marketed, HSV 1 Immunoblot test. One hundred and ninety-eight (82%) of the specimens were obtained during the first trimester, nineteen (8%) during the second trimester and twenty-five (10%) during the third trimester of pregnancy. The results of this comparative test are shown in Table 5 below.

Table 5.

Sensitivity and Specificity of the SeraQuest HSV 1 Type Specific IqG Test in Comparison with a Commercial HSV 1 Immunoblot Test for 242 Expectant Mothers Whose Sera Had Been Submitted for Herpes Simplex Virus Serology.

Seraquest HSV 1 Type Specific IgG Result				% Agreement*		95% C.I.*
HSV 1 ImmunoblotResult	Positive	Equivocal	Negative
Positive	111	3	5	Sensitivity	93.3	87.3 to 96.6
Negative	13	0	110	Specificity	89.4	87.1 to 93.7

CDC Panel Results

The following information was obtained with the Centers for Disease Control and Prevention (CDC) serum panel for HSV serology assays, which was tested in-house by the SeraQuest HSV Type 1 Specific IgG test. The results are presented as a means to convey further information on the performance of this assay with a masked, well characterized serum panel. This does not

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imply an endorsement by the CDC.

Percent Agreement with the CDC Panel

The panel consists of 46 HSV-1 IgG positive and 54 HSV-1 IgG negative samples. The results of this study are shown below in Table 7.

Table 6. SeraQuest HSV Type 1 Specific IgG Results Obtained with the CDC HSV Panel of 100 Sera

		SeraQuest HSV Type 1 Specific IgG
		Positive	Equivocal	Negative	Total
CDC HSV 1 Result	Positive	42	2	2	46
	Negative	0	1	53	54
	Total	42	3	55	100

Conclusion

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

Regulation Number and Section

§ 866.3305 Herpes simplex virus serological assays.

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