Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a standard immunoassay technology and does not mention any AI or ML components in the device description, intended use, or performance studies.

Therapeutic

No
This device is an in vitro diagnostic (IVD) assay designed to detect antibodies to HSV-2, aiding in the presumptive diagnosis of HSV infection. It is not intended to treat or prevent a disease, but rather to diagnose it.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the device is "for in vitro diagnostic use in the qualitative determination of IgG antibodies to herpes simplex virus type 2 (HSV-2)" and "is indicated for testing sexually active adults or expectant mothers for aiding in the presumptive diagnosis of HSV infection." These phrases clearly indicate its role in diagnosing a medical condition.

SaMD (Software as a Medical Device)

No

The device is an in vitro diagnostic assay that uses chemical reagents and an automated system (ADVIA Centaur systems) to perform a physical test on biological specimens. It is not solely software.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use / Indications for Use: The very first sentence explicitly states "The ADVIA Centaur® Herpes-2 IgG (HSV2) assay is for in vitro diagnostic use". It also describes its use in determining the presence of antibodies in human serum and plasma, which are biological samples tested outside of the body.
Device Description: The description details a laboratory assay that analyzes biological specimens (serum and plasma) to detect a specific analyte (IgG antibodies to HSV-2). This is a hallmark of an in vitro diagnostic device.
Clinical Study: The document describes a clinical study where the device was used to test human specimens and the results were compared to reference methods. This is a standard process for evaluating the performance of IVDs.
Key Metrics: The document provides performance metrics like sensitivity and specificity, which are crucial for evaluating the accuracy and reliability of an IVD.
Predicate Device(s): The mention of a predicate device (K000238 Focus HerpeSelect 1 and 2 Immunoblot IgG) further confirms its classification as an IVD, as predicate devices are used for comparison in the regulatory submission process for new IVDs.

All of these points strongly indicate that the ADVIA Centaur® Herpes-2 IgG (HSV2) assay is an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The ADVIA Centaur® Herpes-2 IgG (HSV2) assay is for in vitro diagnostic use in the qualitative determination of IgG antibodies to herpes simplex virus type 2 (HSV-2) in human serum and plasma (EDTA and lithium heparin) using the ADVIA Centaur systems. The test is indicated for testing sexually active adults or expectant mothers for aiding in the presumptive diagnosis of HSV infection. The predictive value of a positive or negative result depends on the prevalence of HSV-2 infection in the population and the pre-test likelihood of HSV-2 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 and plasma (EDTA and lithium heparin).

Product codes

MYF

Device Description

The ADVIA Centaur Herpes-2 IgG (HSV2) assay is a fully automated two-step sandwich immunoassay using indirect chemiluminometric technology. The specimen is incubated with the Solid Phase, which contains HSV-2-specific recombinant-gG2 antigen. Antigen-antibody complexes will form if anti-HSV-2 antibody is present in the specimen. The Lite Reagent contains monoclonal anti-human IgG labeled with acridinium ester, and is used to detect HSV-2 IgG in the specimen.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Sexually active adults or expectant mothers. Not established for pediatric patients.

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

Not Found

Summary of Performance Studies

Precision Study:

Study Type: Precision study according to CLSI EP05-A3.
Sample Size: 6 samples prepared at different levels (Negative and Positive Controls, 6 samples)
Methodology: Each material was tested in duplicate, twice a day for 20 days, for a total of 80 replicates per level.
Key Results:
- Negative Control (Plasma), N=80, Mean=0.29, Repeatability CV=NA, Within-Lab CV=NA
- Positive Control (Plasma), N=80, Mean=3.09, Repeatability CV=2.3%, Within-Lab CV=6.5%
- Serum 1, N=80, Mean=0.36, Repeatability CV=NA, Within-Lab CV=NA
- Plasma 2, N=80, Mean=0.63, Repeatability CV=2.4%, Within-Lab CV=5.9%
- Serum 3, N=80, Mean=1.08, Repeatability CV=4.3%, Within-Lab CV=7.2%
- Serum 4, N=80, Mean=2.58, Repeatability CV=3.6%, Within-Lab CV=7.6%
- Serum 5, N=80, Mean=5.29, Repeatability CV=1.7%, Within-Lab CV=6.4%
- Serum 6, N=80, Mean=7.62, Repeatability CV=2.2%, Within-Lab CV=6.1%
Conclusion: Results met the design requirements for repeatability and within-lab precision.

Sample Matrix Study:

Study Type: Comparison of different sample matrices.
Sample Size: 68 sets of matched samples (serum, serum separator tube (SST), EDTA plasma and lithium heparin plasma) from commercial sources.
Methodology: Samples analyzed in duplicate in randomized order using one reagent lot. Deming regression and linear regression were used for comparison.
Key Results:
- Serum Separator Tube (y) vs Serum (x): y = 0.00 + 0.99x, r=0.999
- EDTA Plasma(y) vs Serum (x): y = 0.00 + 0.98 x, r=0.998
- Lithium Heparin Plasma(y) vs Serum (x): y = -0.01 + 0.98 x, r=0.997
Conclusion: Serum Separator Tube, EDTA Plasma, and Lithium Heparin Plasma are equivalent matrices to Serum.

Panel Studies:

Study Type: Evaluation with commercial sample panels.
Sample Size: ToRCH-mixed Zeptometrix (24 characterized HSV samples), CDC panel (100 blind characterized HSV samples).
Methodology: Samples evaluated with the ADVIA Centaur Herpes-2 IgG assay on ADVIA Centaur XP instrument.
Key Results:
- ToRCH-mixed Zeptometrix panel: 100% total agreement with reference assay 1.
- CDC panel: 100% total agreement in concordance with the results provided by the CDC.

Interferences Study:

Study Type: Assessment of potential interference from common compounds.
Methodology: Interfering substances tested at indicated levels as described in CLSI Document EP7-A2, in three levels of samples (high negative, low positive, and positive) with at least one reagent lot.
Key Results: No interference (≤ 10% change in results) for the ADVIA Centaur Herpes-2 IgG up to the following concentrations: Biotin (3500 ng/mL), Hemoglobin (500 mg/dL), Bilirubin complex (40 mg/dL), Bilirubin free (40 mg/dL), Hypoproteinemia (3 g/dL), Hyperproteinemia (12 g/dL), Lipemia (1000 mg/dL), Cholesterol (400 mg/dL).

Cross-Reactivity Study:

Study Type: Evaluation for potential cross-reactivity with other viral antibodies, disease-state specimens, and other populations.
Sample Size: 522 specimens.
Methodology: HSV-2 IgG status verified using a Comparative Assay; equivocal specimens resolved by Western Blot testing at a reference laboratory.
Key Results: Total percent agreement for clinical categories was 96.9% (506/522).

Multisite Reproducibility Study:

Study Type: Reproducibility evaluated according to CLSI protocol EP5-A3.25.
Sample Size: Not explicitly stated, but involved a sample panel composed of Controls (Negative and Positive) and 6 samples prepared at different levels.
Methodology: Conducted using 1 reagent lot at 3 external sites on the ADVIA Centaur XP instrument. Protocol run over 5 days, 2 runs per day, and 3 replicates per run for samples and controls, pooled results from 3 sites.
Key Results: All results for reproducibility CV were within the design requirement of ≤ 15% for specimens with ≥ 0.80 Index.
- Negative control, N=90, Mean=0.31, Reproducibility %CV=7.9%
- Positive control, N=90, Mean=3.27, Reproducibility %CV=3.2%
- Serum 1, N=90, Mean=0.36, Reproducibility %CV=7.1%
- Plasma 2, N=90, Mean=0.64, Reproducibility %CV=3.4%
- Serum 3, N=90, Mean=1.07, Reproducibility %CV=5.2%
- Serum 4, N=90, Mean=2.47, Reproducibility %CV=7.0%
- Serum 5, N=90, Mean=5.24, Reproducibility %CV=8.2%
- Serum 6, N=90, Mean=7.87, Reproducibility %CV=4.6%

Clinical Study:

Study Type: Multicenter clinical study comparing the reference device with ADVIA Centaur Herpes-2 IgG.
Sample Size: 864 specimens (≥ 18 years of age), including 274 pregnant women.
Data Source: Specimens collected within the United States and tested at 3 independent external laboratories.
Methodology: Sensitivity and specificity determined by comparing performance to a commercially available anti-HSV-2 IgG immunoblot method (Comparative Assay) and a validated Western Blot reference confirmatory test (University of Washington, Seattle). Samples analyzed in singlicate. Equivocal results were resolved by Western Blot.
Key Results (Intended Use Population):
- Overall agreement: 97.6% (843/864) with a 95% CI of 96.3%-98.4%.
- Sensitivity: 95.3% (245/257), 95% CI: 92.0%-97.3%.
- Specificity: 98.5% (598/607), 95% CI: 97.2%-99.2%.
Key Results (Pregnant women population):
- Sensitivity: 100.0% (34/34) with a 95% CI of 89.9%—100.0%.
- Specificity: 98.3% (236/240) with a 95% CI of 95.8%–99.4%.

Key Metrics

Overall agreement (Intended Use Population): 97.6% (843/864) with a 95% confidence interval (CI) of 96.3%-98.4%.
Sensitivity (Intended Use Population): 95.3% (245/257), with a 95% confidence interval (CI) of 92.0%-97.3%.
Specificity (Intended Use Population): 98.5% (598/607), with a 95% confidence interval (CI) of 97.2%-99.2%.
Sensitivity (Pregnant Women Population): 100.0% (34/34) with a 95% confidence interval of 89.9%—100.0%.
Specificity (Pregnant Women Population): 98.3% (236/240) with a 95% confidence interval of 95.8%–99.4%.

Predicate Device(s)

K000238 (Focus HerpeSelect 1 and 2 Immunoblot IgG)

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.3305 Herpes simplex virus serological assays.

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

Summary

0

Image /page/0/Picture/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 a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

August 23, 2018

Biokit, S.A. Joan Guixe QA & RA Director Can Male. S/N Llica d'Amunt, 08186 Barcelona, Spain

Re: K181334

Trade/Device Name: ADVIA Centaur Herpes-2 IgG Regulation Number: 21 CFR 866.3305 Regulation Name: Herpes simplex virus serological assays Regulatory Class: Class II Product Code: MYF Dated: May 17, 2018 Received: May 25, 2018

Dear Joan Guixe:

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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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

1

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) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.html; good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; 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 comprehensive regulatory information about mediation-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 (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely.

Steven R. Gitterman -S for

Uwe Scherf, 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) K183334

Device Name ADVIA Centaur Herpes-2 IgG

Indications for Use (Describe)

The ADVIA Centaur® Herpes-2 IgG (HSV2) assay is for in vitro diagnostic use in the qualitative determination of IgG antibodies to herpes simplex virus type 2 (HSV-2) in human serum and plasma (EDTA and lithium heparin) using the ADVIA Centaur systems. The test is indicated for testing sexually active adults or expectant mothers for aiding in the presumptive diagnosis of HSV infection. The predictive or negative result depends on the prevalence of HSV-2 infection in the population and the pre-test likelihood of HSV-2 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, pediatic patients or matrices other than human serum and plasma (EDTA and lithium heparin).

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)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

3

Image /page/3/Picture/0 description: The image shows the logo for Biokit, a Werfen Company. The logo consists of a stylized blue symbol resembling two overlapping letter 'V' shapes. To the right of the symbol is the word "Biokit" in gray, and below that is the text "A Werfen Company" in a smaller font size.

510(K) SUMMARY

This 510(k) Summary is being submitted in accordance with the requirements of the Safe Medical Device Act of 1990 and 21 CFR 807.92.

| 1. Submitter's Information | Biokit, S.A.
Can Malé S/N
Lliçà d'Amunt 08186
Barcelona (Spain) |

----------------------------	--------------------------------------------------------------------------

2. Contact Person	Joan Guixer, QA & RA Director
	Phone: +34 93 860 90 00 / +34 657 88 33 47
	Email: jguixer@biokit.com

3. Preparation Date	May 17th, 2018
---------------------	----------------

4. Device Trade Name	ADVIA Centaur Herpes-2 IgG
----------------------	----------------------------

5. Regulatory Information	Regulation Number	21 CFR 866.3305
	Regulation Description	Herpes simplex virus
serological assays
	Classification	Class II Special Controls
	Product Code	MYF
	Classification Panel	Microbiology

6. Predicate Device	K000238 (Focus HerpeSelect 1 and 2 Immunoblot IgG)
---------------------	----------------------------------------------------

4

Image /page/4/Picture/0 description: The image shows the logo for Biokit, a Werfen Company. The logo consists of a blue abstract shape resembling two overlapping triangles on the left. To the right of the shape is the word "Biokit" in a gray, sans-serif font. Below the word "Biokit" is the text "A Werfen Company" in a smaller, gray, sans-serif font.

| 7. Indications for Use /
Intended Use | The ADVIA Centaur® Herpes-2 IgG (HSV2) assay is for in
vitro diagnostic use in the qualitative determination of
IgG antibodies to herpes simplex virus type 2 (HSV-2) in
human serum and plasma (EDTA and lithium heparin)
using the ADVIA Centaur systems. The test is indicated
for testing sexually active adults or expectant mothers
for aiding in the presumptive diagnosis of HSV
infection. The predictive value of a positive or negative
result depends on the prevalence of HSV-2 infection in
the population and the pre-test likelihood of HSV-2
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
and plasma (EDTA and lithium heparin). |

| 8. Device Description | The ADVIA Centaur Herpes-2 IgG (HSV2) assay is a fully
automated two-step sandwich immunoassay using
indirect chemiluminometric technology. The specimen
is incubated with the Solid Phase, which contains HSV-
2-specific recombinant-gG2 antigen. Antigen-antibody
complexes will form if anti-HSV-2 antibody is present in
the specimen. The Lite Reagent contains monoclonal
anti-human IgG labeled with acridinium ester, and is
used to detect HSV-2 IgG in the specimen. |

-----------------------

---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

| COMPARISON PREDICATE | | | Manufacturer | Focus Diagnostics
Cypress, CA 90630 -USA | Siemens Heathcare
Diagnostics Inc.
511 Benedict Avenue,
Tarrytown, NY 10591- USA |
|----------------------|---------------------------------------------|----------------------------|--------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Item | Predicate | New Device | Similarities | | |
| Trade Names | Focus HerpeSelect 1 and 2
Immunoblot IgG | ADVIA Centaur Herpes-2 IgG | Intended use | Focus Diagnostics HerpeSelect 1
and 2 Immunoblot IgG test is
intended for qualitatively detecting
the presence or absence of human
IgG class antibodies to HSV-1 and
HSV-2 in human sera. The test is
indicated for testing sexually active
adults or expectant mothers for
aiding in the presumptive diagnosis
of HSV-1 and HSV-2 infection. The
predictive value of a positive or
negative result depends on the
population's prevalence and the
pretest likelihood of HSV- 1 and
HSV-2 infection. The performance
of this assay has not been
established for use in a pediatric
population, for neonatal screening,
for testing of
immunocompromised patients, for
use by a point of care facility or for
use with automated equipment | The ADVIA Centaur® Herpes-
2 IgG (HSV2) assay is for in
vitro diagnostic use in the
qualitative determination of
lgG antibodies to herpes
simplex virus type 2 (HSV-2)
in human serum and plasma
(EDTA and lithium heparin)
using the ADVIA Centaur
systems. The test is indicated
for testing sexually active
adults or expectant mothers
for aiding in the presumptive
diagnosis of HSV infection.
The predictive value of a
positive or negative result
depends on the prevalence of
HSV-2 infection in the
population and the pre-test
likelihood of HSV-2 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 and plasma (EDTA and
lithium heparin). |
| 510K nº | K000238 | K183334 | Measurand | To detect the presence or absence
of human IgG class antibodies to
HSV-1 and HSV-2 | To detect IgG antibodies to
herpes simplex virus type 2
(HSV-2). |
| Regulation Section | 21 CFR 866.3305 | Same | | | |

5

Image /page/5/Picture/0 description: The image shows the logo for Biokit, a Werfen company. The logo consists of a stylized blue "W" shape on the left, followed by the word "Biokit" in gray. Below "Biokit" is the text "A Werfen Company" in a smaller font size, also in gray. The logo is simple and modern, with a focus on the company name and affiliation.

6

Image /page/6/Picture/0 description: The image shows the logo for Biokit. The logo consists of a blue icon on the left and the word "Biokit" in gray on the right. Below the word "Biokit" is the phrase "A Werfen Company" in a smaller font.

Product Code	LGC	MYF
Classification	Class II Special Controls	Same
Assay Type	Qualitative	Same
Differences
Technology	Nitrocellulose immunoblot	Chemiluminescent
immunoassay
Sample type	Human serum	Human serum and
plasma (EDTA and lithium
heparin)

9. Performance Summary

Precision

A precision study was performed according to CLSI EP05-A3 , using the Negative and Positive Controls as well as 6 samples prepared at different levels in the assay range. Each material was tested in duplicate, twice a day for 20 days, for a total of 80 replicates per level.

The ADVIA Centaur HSV2 assay is designed to have the following repeatability and withinlab precision requirements:

	Design Requirement
ADVIA Centaur HSV2 Level (Index)	Repeatability
% CV	Within-Lab
% CV
6.00	≤ 5.0%	≤ 7.0%

೩ NA = not applicable

The results for this study are presented below.

7

Image /page/7/Picture/0 description: The image shows the logo for Biokit, a Werfen Company. The logo consists of a blue abstract graphic to the left of the word "Biokit" in gray. Below the word "Biokit" is the text "A Werfen Company" in a smaller, gray font.

| Specimen Type | N | Mean
(Index) | Repeatability | | Within-Lab | |
|------------------------------|----|-----------------|---------------|-----------|------------|-----------|
| | | | SD | CV
(%) | SD | CV
(%) |
| Negative Control
(Plasma) | 80 | 0.29 | 0.01 | NAa | 0.02 | NAa |
| Positive Control
(Plasma) | 80 | 3.09 | 0.07 | 2.3 | 0.20 | 6.5 |
| Serum 1 | 80 | 0.36 | 0.01 | NAa | 0.02 | NAa |
| Plasma 2 | 80 | 0.63 | 0.02 | 2.4 | 0.04 | 5.9 |
| Serum 3 | 80 | 1.08 | 0.05 | 4.3 | 0.08 | 7.2 |
| Serum 4 | 80 | 2.58 | 0.09 | 3.6 | 0.20 | 7.6 |
| Serum 5 | 80 | 5.29 | 0.09 | 1.7 | 0.34 | 6.4 |
| Serum 6 | 80 | 7.62 | 0.17 | 2.2 | 0.47 | 6.1 |

Precision results for the HSV-2 Assay

NAª= Not applicable

Sample matrix

This study was performed on one ADVIA Centaur XP instrument, using 68 sets of matched samples of different matrixes (serum, serum separator tube (SST), EDTA plasma and lithium heparin plasma) from commercial sources.

The samples were analyzed in duplicate in randomized order using one reagent lot.

. Comparing Serum Separator Tube (y) vs Serum (x): (Deming regression) y = 0.00 + 0.99x, r=0.999, sample range for both matrixes 0.02-9.59 Index Value and 0.02-9.49 Index Value respectively. Note - correlation coefficient is calculated using linear regression.
Comparing EDTA Plasma(y) vs Serum (x): (Deming regression) y= 0.00 + 0.98 x, ● r=0.998, sample range for both matrixes 0.02-9.59 Index Value and 0.02-9.34 Index Value respectively. Note - correlation coefficient is calculated using linear regression.
. Comparing Lithium Heparin Plasma(y) vs Serum (x): (Deming regression) y= -0.01 + 0.98 x, r=0.997, sample range for both matrixes 0.02-9.59 Index Value and 0.01-9.23 Index Value respectively. Note - correlation coefficient is calculated using linear regression.

The results supports that Serum Separator Tube is equivalent matrix to Serum, EDTA Plasma is equivalent matrix to Serum and Lithium Heparin Plasma is an equivalent matrix to Serum.

8

Image /page/8/Picture/0 description: The image contains the logo for Biokit, a Werfen Company. The logo consists of a blue abstract graphic to the left of the text "Biokit". Below the text "Biokit" is the text "A Werfen Company" in a smaller font size. The logo is simple and professional in appearance.

Panels

The commercial sample panels ToRCH-mixed Zeptometrix and CDC panel were analysed.

The ToRCH-mixed Zeptometrix panel included 24 characterized HSV samples. All of the samples were evaluated with the ADVIA Centaur Herpes-2 IgG assay on ADVIA Centaur XP instrument and 100% total agreement was observed with reference assay 1.

The CDC panel included 100 blind characterized HSV samples. All of the samples were evaluated with the ADVIA Centaur Herpe-2 IgG assay on ADVIA Centaur XP instrument and 100% total agreement was observed in concordance with the results provided by the CDC.

Interferences

Potential interference in the ADVIA Centaur HSV2 assay from the compounds listed below is designed to be ≤ 10%. Interfering substances at the levels indicated were tested as described in CLSI Document EP7-A2. The testing was done in three levels of samples in the assay range (high negative, low positive and positive samples) with at least one reagent lot on ADVIA Centaur XP instrument.

Testing confirmed no interference (Demonstrate ≤ 10% change in results) for the ADVIA Centaur Herpes-2 IgG up to the following concentrations:

Serum specimens that are...	Demonstrate ≤10% change in results up to...
Biotin	3500 ng/mL
Hemoglobin	500 mg/dL
Bilirubin complex	40 mg/dL
Bilirubin free	40 mg/dL
Hypoproteinemia	3 g/dL
Hyperproteinemia	12 g/dL
Lipemia	1000 mg/dL
Cholesterol	400 mg/dL

Cross-reactivity

The ADVIA Centaur HSV2 assay was evaluated for potential cross-reactivity other viral antibodies, disease-state specimens, and other populations. The HSV-2 IgG status of each specimen was verified using the Comparative Assay. Repeat equivocal specimens on the Comparative Assay were sent to a reference laboratory for Western Blot testing. Total

9

Image /page/9/Picture/0 description: The image shows the Biokit logo. The logo consists of a blue icon on the left and the word "Biokit" in gray on the right. Below the word "Biokit" is the text "A Werfen Company" in a smaller font size.

percent agreement for the following clinical categories was 96.9% (506/522). The following results were obtained:

| Clinical Category | Number
tested | ADVIA Centaur
HSV2
Reactive | Reference
Method
Positive | |
|----------------------------------------------------|------------------|-----------------------------------|---------------------------------|---------------------|
| Antibody to hepatitis B surface antigen (Anti-HBs) | 10 | 5 | 6 | |
| Anti-gliadin | 10 | 1 | 1 | |
| Anti-nuclear antibodies (ANA) | 10 | 0 | 1 | |
| Candida albicans | 10 | 3 | 3 | |
| Chlamydia trachomatis | 10 | 4 | 4 | |
| Cytomegalovirus (CMV; HHV-5) | 25 | 9 | 11 | |
| Elevated IgG levels | 10 | 8 | 8 | |
| Elevated IgM levels | 10 | 3 | 3 | |
| Epstein-Barr Virus (EBV IgG) | 10 | 3 | 3 | |
| Escherichia coli | 10 | 5 | 5 | |
| Flu vaccine | 10 | 1 | 1 | |
| Gardenerella vaginalis | 10 | 5 | 5 | |
| Hepatitis B surface antigen (HBsAg) | 10 | 4 | 4 | |
| Hepatitis C Virus (HCV) | 10 | 7 | 7 | |
| Herpes simplex virus type 1 (HSV-1) | 117 | 29 | 31 | |
| Heterophile antibodies (EBV) | 10 | 0 | 0 | |
| Human Anti-mouse antibodies (HAMA) | 10 | 4 | 4 | |
| Human herpes virus 6 (HHV-6) | 10 | 2 | 3 | |
| Human herpes virus 8 (HHV-8) | 10 | 5 | 5 | |
| Human immunodeficiency virus (HIV) | 10 | 9 | 9 | |
| Human Papillomavirus (HPV) | 25 | 3 | 5 | |
| | Number
tested | ADVIA Centaur
HSV2 | | Reference
Method |
| Clinical Category | | Reactive | Positive | |
| Multiple myeloma | 10 | 3 | 3 | |
| Neisseria gonorrhoeae | 10 | 4 | 4 | |
| Parvovirus B19 | 10 | 0 | 0 | |
| Rheumatoid factor (RF) | 10 | 5 | 5 | |
| Rubella IgG | 10 | 2 | 2 | |
| Syphilis | 10 | 7 | 7 | |
| Systemic lupus erythematosus (SLE) | 10 | 1 | 0 | |
| Toxoplasma IgG | 95 | 26 | 30 | |
| Varicella Zoster virus (HHV-3) | 10 | 1 | 1 | |
| Total Samples | 522 | 159 | 171 | |

Cross Reactivity Study Performance Results

10

Biokit
A Werfen Company

Multisite reproducibility

Reproducibility was evaluated according to the CLSI protocol EP5-A3.25. The ADVIA Centaur HSV2 assay is designed to have reproducibility precision requirements of ≤ 15% for specimens with ≥ 0.80 Index.

A reproducibility study was conducted using 1 reagent lot at 3 external sites on the ADVIA Centaur XP instrument, using a sample panel blinded and randomized that was composed by the Controls (Negative and Positive) and 6 samples prepared at different levels in the assay range.

The protocol was run over 5 days, 2 runs per day, and 3 replicates per run for the sample pools, and for the negative and positive control materials. The data in the following table represents pooled results from the 3 sites.

The results are presented in the table below.

| Specimen
type | N | Mean
(Index) | Repeatability | | Between-Run | | Between-Day | | Between-Site | | Reproducibility | |
|---------------------|----|-----------------|---------------|------|-------------|------|-------------|------|--------------|------|-----------------|------|
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Negative
control | 90 | 0.31 | 0.01 | 4.5% | 0.00 | 1.1% | 0.01 | 1.8% | 0.02 | 6.2% | 0.02 | 7.9% |

Multisite Precision Study Repeatability

11

Image /page/11/Picture/0 description: The image contains the logo for Biokit. The logo consists of a blue icon on the left and the word "Biokit" in gray on the right. The icon is made up of two overlapping shapes, one darker blue and one lighter blue. The word "Biokit" is in a sans-serif font.

A Werfen Company

| Specimen
type | N | Mean
(Index) | Repeatability | | Between-Run | | Between-Day | | Between-Site | | Reproducibility | |
|------------------|----|-----------------|---------------|------|-------------|------|-------------|------|--------------|------|-----------------|------|
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Positive control | 90 | 3.27 | 0.06 | 1.9% | 0.04 | 1.2% | 0.04 | 1.4% | 0.06 | 1.8% | 0.10 | 3.2% |
| Serum 1 | 90 | 0.36 | 0.01 | 2.3% | 0.01 | 3.6% | 0.01 | 3.1% | 0.02 | 4.8% | 0.03 | 7.1% |
| Plasma 2 | 90 | 0.64 | 0.01 | 1.8% | 0.01 | 1.1% | 0.02 | 2.7% | 0.00 | 0.0% | 0.02 | 3.4% |
| Serum 3 | 90 | 1.07 | 0.02 | 1.8% | 0.03 | 2.8% | 0.03 | 2.9% | 0.03 | 2.8% | 0.06 | 5.2% |
| Serum 4 | 90 | 2.47 | 0.08 | 3.1% | 0.06 | 2.5% | 0.05 | 1.8% | 0.13 | 5.4% | 0.17 | 7.0% |
| Serum 5 | 90 | 5.24 | 0.34 | 6.4% | 0.12 | 2.3% | 0.11 | 2.2% | 0.21 | 4.0% | 0.43 | 8.2% |
| Serum 6 | 90 | 7.87 | 0.14 | 1.8% | 0.18 | 2.2% | 0.20 | 2.5% | 0.20 | 2.5% | 0.36 | 4.6% |

Clinical study

A multicenter clinical study to compare the reference device with ADVIA Centaur Herpes-2 IgG was performed. Sensitivity and specificity were determined by comparing the performance of the ADVIA Centaur HSV2 assay to a commercially available anti-HSV-2 IgG immunoblot method (Comparative Assay) and a validated Western Blot reference confirmatory test (University of Washington, Seattle). A total of 864 specimens (≥ 18 years of age), including specimens from 274 pregnant women, were collected within the United States and tested at 3 independent external laboratories. Of these specimens, 254 were reactive, and 610 were nonreactive with the ADVIA Centaur HSV2 assay. The overall agreement was 97.6% (843/864) with a 95% confidence interval (CI) of 96.3%-98.4%. Samples were analyzed in singlicate.

Of the 864 specimens tested by the Comparative Assay, 22 were equivocal and were further tested by the Western Blot test. After Western Blot testing, 20 of the 22 specimens were resolved to be negative and 2 remained equivocal.

| Intended use population | | Comparative Assay
Equivocal results have been resolved by Western Blot (WB) | Positive | Negative | Equivocal | Total |
|-------------------------|-------------|--------------------------------------------------------------------------------|----------|----------|-----------|-------|
| ADVIA Centaur HSV2 | Reactive | | 245 | 9 | 0 | 254 |
| | Nonreactive | | 10 | 598 | 2 | 610 |
| | Total | | 255 | 607 | 2 | 864 |

The results obtained for the intended use population are presented below

The sensitivity of the ADVIA Centaur HSV2 assay was 95.3% (245/257), with a 95% confidence interval (CI) of 92.0%-97.3%.

The specificity of the ADVIA Centaur HSV2 assay was 98.5% (598/607), with a 95% confidence interval (CI) of 97.2%-99.2%.

12

Image /page/12/Picture/0 description: The image contains the logo for Biokit, a Werfen company. The logo consists of a stylized "W" made up of three blue parallelograms, followed by the company name "Biokit" in gray text. Below the company name, the text "A Werfen Company" is written in a smaller, lighter gray font.

Sensitivity and specificity were determined for the pregnant population by comparing the performance of the ADVIA Centaur HSV2 assay to the Comparative Assay and Western Blot confirmatory method. The results of pregnant women samples are presented below:

| Pregnant women | Comparative assay
Equivocal results have been resolved by Western Blot (WB) | | | | |
|--------------------|--------------------------------------------------------------------------------|----------|-----------|-------|-----|
| | Positive | Negative | Equivocal | Total | |
| ADVIA Centaur HSV2 | Reactive | 34 | 4 | 0 | 38 |
| | Nonreactive | 0 | 236 | 0 | 236 |
| | Total | 34 | 240 | 0 | 274 |

Pregnant women Study Contingency (WB resolution) table

The sensitivity of the ADVIA Centaur HSV2 assay was 100.0% (34/34) with a 95% confidence interval of 89.9%—100.0%.

The specificity of the ADVIA Centaur HSV2 assay was 98.3% (236/240) with a 95% confidence interval of 95.8%–99.4%.

10. Stability

The onboard stability of the ADVIA Centaur Herpes-2 IgG reagents is 60 days with a calibration interval of 28 days. The onboard stability of the ADVIA Centaur Herpes-2 IgG Calibrators is 8 hours. The opened vial stability of the ADVIA Centaur Herpes-2 IgG Calibrators is 65 days. Unopened reagents and calibrators are stable until the date printed on the box label when stored at 2-8°C.

11. Conclusion

The analytical and clinical study results demonstrate that the ADVIA Centaur Herpes-2 IgG are substantially equivalent to the predicate device, Focus HerpeSelect 1 and 2 Immunoblot IgG (FDA cleared under K000238), and that the assay is safe and effective for its labeled intended use.