The Zeus Scientific, Inc. AtheNA Multi-Lyte® ToRCH IgG Plus Test System is intended for the qualitative detection of specific human IgG class antibodies to Toxoplasma gondii (T.gondii), Rubella, Cytomegalovirus (CMV) and HSV 1 & 2 in human serum. The results of this assay are intended to be used as an aid in the assessment of serological status to Toxoplasma gondii, Rubella and CMV. For HSV 1 and HSV 2, the test is indicated for sexually active adults and expectant mothers, as an aid for presumptively diagnosing Herpes Simplex 1 and Herpes Simplex 2.

The test is not intended for use in screening blood or plasma donors.

The performance of this assay has not been established for use in a pediatric population, neonatal screening, immunocompromised or immunosuppressed patients or for use at point of care facilities.

Device Description

The Zeus Scientific, Inc. AtheNA Multi-Lyte® ToRCH IgG Plus Test System is a multiplex immunoassav intended for the simultaneous qualitative detection and differentiation of specific human IgG class antibodies to Toxoplasma gondii, Rubella, Cytomegalovirus (CMV), Herpes Simplex 1 (HSV-1) and Herpes Simplex 2 (HSV-2) in human serum. The results of this assay are intended to be used as an aid in the assessment of a patient's serological status to infection with Toxoplasma gondii, Rubella, CMV, HSV 1 and HSV 2 and in the determination of immune status of individuals including pregnant women. The test system is comprised of the AtheNA Multi-Lyte test kit, software and the Luminex Corp instrument.

The AtheNA Multi-Lyte ToRCH IgG Plus Test System provides the following components:

Reactive Reagents:

All reactive reagents contain sodium azide as a preservative at a concentration of 0.1% (w/v).

Multiplexed bead suspension 1. Ready to use, 5.5 mL bottle. The suspension contains separate distinguishable 5.6 micron polystyrene beads that are conjugated with:
. Toxo grade 2 antigen
. Rubella K2S grade antigen
CMV grade 2
. HSV-1 type-specific recombinant gG-1 protein antigen
. HSV-2 gG-2 type-specific recombinant gG-2 protein antigen

The bead mix also contains one bead set designed to detect non-specific antibodies in the patient sample (if present) and four separate bead sets used for assay calibration.

Conjugate: Phycoerythrin conjugated goat anti-human IgG (y chain specific). Ready to use, 15 mL amber bottle.
Human positive serum control 1. One, 0.2 mL vial.
Human positive serum control 2. One, 0.2 mL vial.
Human negative serum control. One, 0.2 mL vial.
SAVe Diluent®. One 50 mL bottle containing phosphate-buffered-saline. Ready to use. NOTE, the sample diluent will change color in the presence of serum.
Wash Buffer Concentrate: dilute 1 part concentrate + 9 parts deionized or distilled water. One bottle containing 10 X concentrate of phosphate buffered saline.

Non-Reactive Reagents

One, 96-well filtration plate for rinsing the microspheres
Data Labels: One label is adhered to the inside lid of the kit box and a second label is inside the kit box.
Package Insert providing instructions for use
Calibration CD: a compact disc that includes all lot-specific kit calibration values required for specimen analysis and assay quality control

AI/ML Overview

Here's a breakdown of the acceptance criteria and the study that proves the device meets them, based on the provided text:

Device: AtheNA Multi-Lyte® ToRCH IgG Plus Test System
Submission Purpose: Simultaneous qualitative detection and differentiation of specific human IgG class antibodies to Toxoplasma gondii (Toxo), Rubella, Cytomegalovirus (CMV), and Herpes Simplex 1 (HSV-1) and Herpes Simplex 2 (HSV-2). The results aid in assessing serological status for Toxo, Rubella, and CMV, and for presumptively diagnosing HSV-1 and HSV-2 in sexually active adults and expectant mothers.

1. Table of Acceptance Criteria and Reported Device Performance

The document doesn't explicitly state "acceptance criteria" as a single, consolidated list with numerical targets for each performance metric (e.g., "Sensitivity must be >95%"). Instead, it presents the results of comparative studies against predicate devices and CDC panels. The implicit acceptance criterion is that the device demonstrates performance substantially equivalent to or in agreement with these established methods.

The table below summarizes the reported device performance from the clinical studies for various analytes and populations, indicating successful agreement or high percentage agreement where applicable. For Toxoplasma, Rubella, and CMV in the "Individuals Undergoing ToRCH Antibody Assessment," the columns refer to "Positive Percent Agreement (PPA)" which represents sensitivity and "Negative Percent Agreement (NPA)" which represents specificity. For HSV-1 and HSV-2 in "Sexually Active Adults", it uses "Sensitivity" and "Specificity." For the CDC panels, it also uses "PPA" and "NPA" against the CDC results.

Analyte (Population)	Acceptance Criteria (Implicit)	Reported Device Performance
Individuals Undergoing ToRCH Antibody Assessment
Toxoplasma (Positive)	High agreement with predicate device for positive samples	PPA = 99.3% (136/137) (95% CI: 96.0% - 100%)
Toxoplasma (Negative)	High agreement with predicate device for negative samples	NPA = 98.0% (450/514) (95% CI: 98.0% - 99.9%) (Typo in original doc for 95.7%)
Rubella (Positive)	High agreement with predicate device for positive samples	PPA = 98.5% (533/541) (95% CI: 97.1% - 99.4%)
Rubella (Negative)	High agreement with predicate device for negative samples	NPA = 87% (60/69) (95% CI: 76.7% - 93.9%)
CMV (Positive)	High agreement with predicate device for positive samples	PPA = 99.6% (450/452) (95% CI: 98.4% - 100%)
CMV (Negative)	High agreement with predicate device for negative samples	NPA = 91.3% (181/197) (95% CI: 87.2% - 95.3%)
Sexually Active Adults
HSV-1 (Positive)	High sensitivity	Sensitivity = 98.6% (418/424) (95% CI: 97.0% - 99.5%)
HSV-1 (Negative)	High specificity	Specificity = 94.6% (163/172) (95% CI: 90.3% - 97.6%)
HSV-2 (Positive)	High sensitivity	Sensitivity = 96.9% (127/131) (95% CI: 92.4% - 98.8%)
HSV-2 (Negative)	High specificity	Specificity = 93.5% (433/463) (95% CI: 90.9% - 95.6%)
Pregnant Women
Toxoplasma (Positive)	High agreement with predicate device for positive samples	PPA = 91.3% (21/23) (95% CI: 72.0% - 98.9%)
Toxoplasma (Negative)	High agreement with predicate device for negative samples	NPA = 95.3% (170/178) (95% CI: 91.3% - 98.0%)
Rubella (Positive)	High agreement with predicate device for positive samples	PPA = 99.0% (194/196) (95% CI: 96.4% - 99.9%)
Rubella (Negative)	High agreement with predicate device for negative samples	NPA = 100.0% (4/4) (95% CI: 47.3% - 100%)
CMV (Positive)	High agreement with predicate device for positive samples	PPA = 98.1% (151/154) (95% CI: 94.4% - 99.6%)
CMV (Negative)	High agreement with predicate device for negative samples	NPA = 100.0% (46/46) (95% CI: 93.7% - 100%)
HSV-1 (Positive)	High agreement with predicate device for positive samples	PPA = 99.3% (137/138) (95% CI: 96.1% - 100%)
HSV-1 (Negative)	High agreement with predicate device for negative samples	NPA = 85.2% (46/54) (95% CI: 72.3% - 93.4%)
HSV-2 (Positive)	High agreement with predicate device for positive samples	PPA = 97.1% (68/70) (95% CI: 90.1% - 99.7%)
HSV-2 (Negative)	High agreement with predicate device for negative samples	NPA = 92.6% (113/122) (95% CI: 86.5% - 96.6%)
Agreement with CDC Panel
Toxoplasma (Positive)	100% agreement with CDC panel positives	PPA = 100.0% (70/70) (95% CI: 95.8% - 100.0%)
Toxoplasma (Negative)	100% agreement with CDC panel negatives	NPA = 100.0% (30/30) (95% CI: 90.5% - 100.0%)
Rubella (Positive)	100% agreement with CDC panel positives	PPA = 100.0% (80/80) (95% CI: 96.3% - 100.0%)
Rubella (Negative)	100% agreement with CDC panel negatives	NPA = 100.0% (20/20) (95% CI: 86.1% - 100.0%)
CMV (Positive)	100% agreement with CDC panel positives	PPA = 100.0% (52/52) (95% CI: 94.4% - 100.0%)
CMV (Negative)	High agreement with CDC panel negatives	NPA = 95.8% (46/48) (95% CI: 90.2% - 100.0%)
HSV-1 (Positive)	100% agreement with CDC panel positives	PPA = 100.0% (50/50) (95% CI: 94.2% - 100.0%)
HSV-1 (Negative)	100% agreement with CDC panel negatives	NPA = 100.0% (50/50) (95% CI: 94.2% - 100.0%)
HSV-2 (Positive)	100% agreement with CDC panel positives	PPA = 100.0% (48/48) (95% CI: 94.0% - 100.0%)
HSV-2 (Negative)	High agreement with CDC panel negatives	NPA = 98.1% (51/52) (95% CI: 94.3% - 100.0%)

2. Sample Size Used for the Test Set and Data Provenance

The evaluation of the AtheNA Multi-Lyte ToRCH IgG Plus Test System involved several studies:

Comparative testing of Intended Use Populations (651 samples):
- Sample Size: 651 unselected samples.
- Data Provenance:
  - 300 samples from a hospital laboratory in the Mid-Atlantic region (United States).
  - 351 samples from a hospital laboratory in the Northeast (United States).
  - The samples were "prospectively collected" individuals undergoing ToRCH antibody assessment. They were described as "frozen remnant serum samples" which suggests they might have been collected over a period and then accessed retrospectively for this study. The phrasing "prospectively collected frozen remnant serum samples" can be ambiguous, but generally, "prospectively collected" refers to data collected specifically for the study. However, using "remnant" suggests they were left over from routine testing.
  - Data was gathered concurrently with the AtheNA Multi-Lyte test system and predicate assays.
Pregnant Women Population (200 samples):
- Sample Size: 200 samples.
- Data Provenance: From expectant mothers, sourced from two serum vendors. The samples were "prospectively collected" and were "frozen remnant serum samples". Tested internally at the manufacturer's lab.
CDC Reference Panels:
- Sample Size:
  - Toxo: 100 samples (70 positive, 30 negative)
  - Rubella: 100 samples (80 positive, 20 negative)
  - CMV: 100 samples (54 positive, 46 negative)
  - HSV-1: 100 samples (50 positive, 50 negative)
  - HSV-2: 100 samples (48 positive, 52 negative)
- Data Provenance: Masked, well-characterized serum panels from the CDC (Centers for Disease Control and Prevention), a US government agency.
HSV-1 & HSV-2 Low Prevalence Population:
- Sample Size: 67 samples.
- Data Provenance: Serum samples from 18 and 19-year-old subjects previously tested for non-sexual infections. Assessed internally at Zeus. Retrospective.
Rubella Retrospective Negative Sample Study:
- Sample Size: 100 samples.
- Data Provenance: Pre-selected banked samples of sera previously tested negative for Rubella antibody by the predicate device. Assessed internally at Zeus. Retrospective.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications

The document does not explicitly mention the number or qualifications of experts used to establish the ground truth for the test sets. Instead, the "ground truth" was established by:

Predicate assays: The results of FDA-cleared predicate ELISA test systems (Toxo IgG ELISA, Rubella IgG ELISA, CMV IgG ELISA, HerpeSelect 1 and 2 Immunoblot IgG for HSV-1/2) were used as the reference standard for the clinical performance studies in general populations and pregnant women. These predicate devices are themselves validated diagnostic tools.
CDC Reference Panels: These are "well-characterized" serum panels which implicitly means their status (positive/negative) for the target analytes has been rigorously established, likely through expert consensus and/or a battery of highly sensitive and specific reference methods, although the details are not provided here.

4. Adjudication Method for the Test Set

The document does not describe any explicit adjudication method (like 2+1 or 3+1) for resolving discrepancies between the AtheNA Multi-Lyte system and the predicate devices or CDC panels.

The results are presented as direct comparisons, and for discrepant cases in the Rubella category, a note mentions qualitative differences (e.g., "4/4 discrepant Rubella samples which tested positive by ELISA had low positive values for AtheNA and high negative values for ELISA, 4/4 discrepant Rubella samples which tested positive by AtheNA and equivocal by ELISA had low positive values for ELISA"). This suggests discrepancy analysis was performed but no formal adjudication by an independent panel of experts is explicitly mentioned for overriding results.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done

No, an MRMC comparative effectiveness study was not done. This device is an automated in vitro diagnostic assay (multiplex microparticle immunoassay) that generates quantitative/qualitative results, not an imaging AI algorithm requiring human interpretation. Therefore, the concept of "human readers improve with AI vs without AI assistance" does not apply in this context.

6. If a Standalone Study (Algorithm Only Without Human-in-the-Loop Performance) Was Done

Yes, the studies reported are essentially standalone performance studies. The AtheNA Multi-Lyte® ToRCH IgG Plus Test System is an automated immunoassay system (test kit, software, and Luminex Corp instrument) which provides qualitative detection of antibodies. Its performance is evaluated directly against predicate devices or reference panels without human interpretation influencing the device's output. The results presented (PPA, NPA, Sensitivity, Specificity) are inherent to the device's diagnostic capability.

7. The Type of Ground Truth Used

The ground truth used for the test sets was primarily based on:

Predicate devices: Established and FDA-cleared immunoassay systems for each analyte (ELISA for Toxo, Rubella, CMV; Immunoblot IgG for HSV-1/2).
CDC Reference Panels: Well-characterized serum panels with known positive or negative status.
Previously tested results: For the low prevalence HSV study and retrospective Rubella study, pre-selected banked samples with known results from predicate devices were used.

These are considered established diagnostic results from reference methods or highly characterized samples, rather than pathology or direct outcomes data from patients.

8. The Sample Size for the Training Set

The document does not explicitly mention a "training set" or its sample size for the AtheNA Multi-Lyte® ToRCH IgG Plus Test System. As an immunoassay, the device itself is developed and optimized through laboratory procedures, reagent formulation, and analytical validation rather than machine learning training on a dataset. The studies described are performance evaluation studies, akin to a test set, to demonstrate substantial equivalence and establish performance characteristics.

9. How the Ground Truth for the Training Set Was Established

Since a "training set" in the context of machine learning is not applicable here, the method of establishing ground truth for a training set is not provided. The development of an immunoassay involves optimizing reagents and reaction conditions, typically through analytical experiments rather than data-driven machine learning training.

Summary

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JUL 1 6 2010

Zeus Scientific Inc. 510K Summary:

AtheNA Multi-Lyte® ToRCH IgG Plus Test System

Administrative Information

1. 510(k): number: K093784
1. Submission Purpose: Intent to market referenced device for the simultaneous qualitative detection and differentiation of specific human IgG class antibodies to Toxoplasma gondii (Toxo), Rubella, Cytomegalovirus (CMV) and Herpes Simplex 1 (HSV-1) and Herpes Simplex 2 (HSV-2).
1. Measurand: Toxoplasma, Rubella, CMV, HSV 1 and HSV 2 IgG antibodies.
1. Type of Test: Multiplex microparticle immunoassay.
Applicant: Zeus Scientific, Inc. (Zeus), PO Box 38, Raritan, NJ 08869 (908)526-3744 5.
1. Proprietary Name: AtheNA Multi-Lyte® ToRCH IgG Plus Test System.
1. Established name: Toxoplasma, Rubella, Cytomegalovirus, Herpes Simplex 1 and Herpes Simplex 2 serological reagents

Regulatory Information

Toxo

Device Classification: Enzyme Linked Immunoabsorbent Assay Regulation Description: Toxoplasma gondii serological reagents Class: Class 2 Product Code: LGD Panel: Microbiology Regulation Number: 866.3780

Rubella

Device Classification: Enzyme Linked Immunoabsorbent Assay Regulation Description: Rubella virus serological reagents Class: Class 2 Product Code: LFX Panel: Microbiology Regulation Number: 866.3510

CMV

Device Classification: Enzyme Linked Immunoabsorbent Assay Regulation Description: Cytomegalovirus serological reagents Class: Class 2 Product Code: LFZ Panel: Microbiology Regulation Number: 866.3175

HSV-1

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Device Classification: Enzyme Linked Immunoabsorbent Assay Regulation Description: Herpes Simplex virus serological reagents Class: Class 2 Product Code: MXJ Panel: Microbiology Regulation Number: 866.3305

HSV-2

Device Classification: Enzyme Linked Immunoabsorbent Assay Regulation Description: Herpes Simplex 2 serological reagents Class: Class 2 Product Code: MYF Panel: Microbiology Regulation Number: 866.3305

Intended Use

The test is not intended for use in screening blood or plasma donors.

The performance of this assay has not been established for use in a pediatric population, neonatal screening, immunocompromised or immunosuppressed patients or for use at point of care facilities.

Device Description

The AtheNA Multi-Lyte ToRCH IgG Plus Test System provides the following components:

Reactive Reagents:

All reactive reagents contain sodium azide as a preservative at a concentration of 0.1% (w/v).

1. Multiplexed bead suspension 1. Ready to use, 5.5 mL bottle. The suspension contains separate distinguishable 5.6 micron polystyrene beads that are conjugated with:
- . Toxo grade 2 antigen

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. Rubella K2S grade antigen
CMV grade 2
. HSV-1 type-specific recombinant gG-1 protein antigen
. HSV-2 gG-2 type-specific recombinant gG-2 protein antigen

The bead mix also contains one bead set designed to detect non-specific antibodies in the patient sample (if present) and four separate bead sets used for assay calibration.

1. Conjugate: Phycoerythrin conjugated goat anti-human IgG (y chain specific). Ready to use, 15 mL amber bottle.
1. Human positive serum control 1. One, 0.2 mL vial.
1. Human positive serum control 2. One, 0.2 mL vial.
1. Human negative serum control. One, 0.2 mL vial.
1. SAVe Diluent®. One 50 mL bottle containing phosphate-buffered-saline. Ready to use. NOTE, the sample diluent will change color in the presence of serum.
1. Wash Buffer Concentrate: dilute 1 part concentrate + 9 parts deionized or distilled water. One bottle containing 10 X concentrate of phosphate buffered saline.

Non-Reactive Reagents

1. One, 96-well filtration plate for rinsing the microspheres
1. Data Labels: One label is adhered to the inside lid of the kit box and a second label is inside the kit box.
1. Package Insert providing instructions for use
1. Calibration CD: a compact disc that includes all lot-specific kit calibration values required for specimen analysis and assay quality control

Substantial Equivalence

Examination of enclosed data indicates that the Zeus Scientific, Inc AtheNA Multi-Lyte ToRCH IgG Plus Test System for the simultaneous detection and differentiation of IgG class antibodies to Toxo, Rubella, CMV, HSV-1 and HSV-2 is substantially equivalent to commercially marketed test systems which have been previously cleared by the FDA for in vitro diagnostic use.

The comparator information for both cleared and investigational analytes are listed:

Toxoplasma

1. Name of Predicate Device: Toxo IgG ELISA Test System
Manufacturer of Predicate Device: Zeus Scientific, Inc. 2.
1. 510(k)891781

Rubella

1. Name of Predicate Device: Rubella IgG ELISA Test System
1. Manufacturer of Predicate Device: Zeus Scientific, Inc.
1. 510(k)891783

CMV

Name of Predicate Device: CMV IgG ELISA Test System 1.
1. Manufacturer of Predicate Device: Zeus Scientific, Inc.

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1. 510(k)924096

HSV-I

Name of Predicate Device: HerpeSelect 1 and 2 Immunoblot IgG 1.
Manufacturer of Predicate Device: Focus Diagnostics 2.
1. 510(k)000238

HSV-2

Name of Predicate Device: HerpeSelect 1 and 2 Immunoblot IgG 1.
1. Manufacturer of Predicate Device: Focus Diagnostics
1. 510(k)000238

Test Principle

The Zeus Scientific, Inc. AtheNA Multi-Lyte ToRCH IgG Plus Test System is designed to detect IgG class antibodies in human sera to Toxo, Rubella, CMV, HSV-2. The test procedure involves four incubation steps:

1. Patient sera are diluted test sera are incubated in a vessel containing a multiplexed mixture of the bead suspension. The multiplexed bead suspension contains a mixture of distinguishable sets of polystyrene microspheres; each set conjugated with Toxoplasma, Rubella. CMV, HSV-1 and HSV-2 antigens. The bead mix also contains one bead set designed to detect nonspecific antibodies in the patient sample (if present) and four separate bead sets used for assay calibration. If present in patient sera, the individual antibodies will bind to the corresponding immobilized antigen bead set. The microspheres are rinsed to remove non-reactive serum proteins.
1. Phycoerythrin-conjugated goat anti-human lgG is added to the vessel and the plate is incubated. The conjugate will react with IgG antibody immobilized on the beads in step 1. The bead suspension is then analyzed by the AtheNA Multi-Lyte® instrument. The bead set(s) are sorted (identified) and the amount of reporter molecule (PE conjugate) is determined for each bead set. Using the Intra-Well Calibration Technology®, internal calibration bead sets are used to to convert raw fluorescence into outcome (units).

Analytical Specificity Interfering Substances

The effect of potential interfering substances on sample results generated using the AtheNA Multi-Lyte test system was evaluated with the following possible interfering substances based on the guidelines established in CLSI EP7-A2: albumin, bilirubin, cholesterol, hemoglobin, triglycerides and intralipids.

The quantity of analyte in each interfering substance is as follows and is based on CLSI EP7-A2:

Bilirubin: 1mg/dL (low), 15 mg/dL (high)

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Albumin: 3.5 g/dL (low), 5 g/dL (high) Cholesterol: 150 mg/dL (low), 250 mg/dL (high) Triglycerides: 150 mg/dL (low), 500 mg/dL (high) Hemoglobin: 20 g/dL (low), 20 g/dL (high) Intralipid: 300 mg/dL (low), 750 mg/dL (high)

Three samples each for Toxo, Rubella, CMV, HSV 1 and 2 IgG were chosen based on their performance on the AtheNA Multi-Lyte test system: positive, borderline and negative. The samples were exposed to the possible interfering substance and tested. All samples showed less than a 20% change in signal with the following exceptions:

	Potential Interfering Substance Spikes Exhibiting Change in Signal Greater than 20%
	Bilirubin		Albumin		Cholesterol		Triglycerides		Hemoglobin		Intralipid
Analyte/Level of Sample	High	Low	High	Low	High	Low	High	Low	High	Low	High	Low
Toxoplasma Positive	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	22%	<20%	<20%
Toxoplasma Borderline	<20%	<20%	-33%	-30%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%
Toxoplasma Negative	<20%	<20%	78%	89%	31%	<20%	<20%	<20%	<20%	22%	78%	<20%
Rubella Positive	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	24%	<20%	<20%
Rubella Borderline	<20%	<20%	<20%	<20%	-27%	<20%	<20%	<20%	<20%	-31%	<20%	<20%
Rubella Negative	50%	50%	<20%	-33%	<20%	-33%	-33%	-33%	<20%	-33%	<20%	<20%
CMV Positive	<20%	<20%	-27%	-25%	-32%	-31%	<20%	<20%	-33%	-35%	<20%	<20%
CMV Borderline	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	31%	24%	<20%
CMV Negative	<20%	25%	<20%	32%	<20%	<20%	<20%	<20%	<20%	25%	<20%	25%
HSV 1 Postive	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	27%	22%	<20%	<20%
HSV 1 Borderline	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%
HSV 1 Negative	<20%	<20%	41%	36%	<20%	<20%	<20%	20%	56%	26%	<20%	<20%
HSV 2 Positive	<20%	<20%	<20%	<20%	27%	<20%	24%	21%	<20%	23%	<20%	<20%
HSV 2 Borderline	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%	<20%
HSV 2 Negative	50%	41%	<20%	<20%	<20%	<20%	<20%	<20%	32%	35%	<20%	<20%

Cross Reactivity

Studies were performed at the manufacturing facility to assess cross reactivity with the Athena Multi-Lyte ToRCH IgG Plus test system using samples that were sero-positive to Measles, Mumps, Rubella, VZV, EBV VCA IgG, EBNA-1, HSV-2, CMV, Syphilis, Toxoplasma and ANA and Rf IgM. Micro-particle and ELISA immunoassay test systems manufactured for commercial distribution were used to determine the sero-positivity of the samples minimally for each possible cross-reactant were tested. The results presented were obtained by testing the analytes against high concentrations of possible cross reactants.

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	AtheNA Multi-Lyte ToRCH IgG Plus Cross Reactivity Summary
Analyte	Toxoplasma	Rubella	CMV	HSV 1	HSV 2
Measles	0/10	0/20	0/10	0/10	0/10
Mumps	0/10	0/20	0/10	0/10	0/10
Rubella	0/10	N/A	0/10	0/10	0/10
VZV	0/10	0/20	0/10	0/10	0/10
VCA IgG	0/10	0/20	0/10	0/10	0/10
EBNA IgG	0/10	0/20	0/10	0/10	0/10
HSV 1	0/10	0/20	0/10	N/A	0/10
HSV 2	0/10	0/20	0/10	0/10	N/A
ANA	0/10	0/4	0/10	0/10	0/10
RF	0/10	0/10	0/10	0/10	0/10
CMV	0/10	0/20	N/A	0/10	0/10
Syphilis	0/10	0/10	0/10	0/10	0/10
Toxoplasma	NA	0/20	0/10	0/10	0/10

Clinical Performance

Clinical Data Generated for Submission

Comparative testing of the Intended Use Populations: AtheNA Multi-Lyte ToRCH IgG Plus 1. versus predicate assays. 851 unselected samples submitted for ToRCH testing were obtained from various serum vendors. The samples were submitted for ToRCH antibody testing, sequentially numbered, de-identified and archived and made available for purchase. After procurement, 300 samples were tested at a hospital laboratory located in the Mid-Atlantic region; 351 samples were tested at a hospital laboratory located in the Northeast. 200 samples were from Expectant Mothers and were tested at the manufacturing site's laboratory. All results are from data generated concurrently using the AtheNA Multi-Lyte ToRCH IgG Plus test system.
1. Comparative testing in Populations other than those for Intended Use: Additional clinical performance was assessed in special populations other than the intended use populations. Performance data was gathered on HSV-1 and HSV-2 low prevalence populations and the relative specificity of Rubella was assessed internally at Zeus using pre-selected banked samples of sera which previously tested negative for Rubella antibody by the predicate device
1. Clinical performance with International Standards and CDC Reference Panels: Performance of the AtheNA Multi-Lyte ToRCH IgG Plus was assessed using the CDC Reference Panels. Traceability and linearity was demonstrated for Rubella using the WHO standard. The performance of Rubella IgG was demonstrated using the CDC low titer standard.
Precision and Reproducibility: Precision of the device was assessed using nine samples. These 4. repeatability studies were performed internally at the manufacturing site's laboratory. Reproducibility was assessed using six samples tested at three sites, two external and one internal.

Expected Results

Observed prevalence was evaluated at three sites in a prospective study including individuals and pregnant women undergoing ToRCH testing.

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1. 651 masked samples prospectively collected from individuals between the ages of <1 and 89 were tested at two external sites. 300 samples were submitted for ToRCH antibody assessment. 351 samples were submitted for testing of one or more of the anlytes in the ToRCH panel. Testng was performed on all five markers on all samples. Results from a subset of 596/651 individuals between the ages of 17 and 69 were used to calculate HSV 1 and HSV 2 prevalence. Site 1, a hospital laboratory located in the Mid-Atlantic region tested 300 samples. Site 2, a hospital laboratory in the Northeast tested 351 samples.
1. 200 masked samples prospectively collected from pregnant women for ToRCH antibody assessment were obtained from two serum vendors. The women ranged in age from 15 to 46.The samples were tested internally at the manufacturer site for all five analytes.

AtheNA Multi-Lyte ToRCH IgG Plus Observed Prevalence in Intended Use Populations

Observed Prevalence in Individuals Undergoing TORCH Antibody AssessmentPrevalence of Analytes in Prospective Samples
		Toxoplasma		Rubella		CMV
Age	Gender	Pos/Total	%Prevalence	Pos/Total	%Prevalence	Pos/Total	%Prevalence
0-9	Male	0/2	0.0%	1/2	50.0%	1/2	50.0%
	Female	1/2	50.0%	1/2	50.0%	1/2	50.0%
10-19	Male	1/3	33.3%	3/3	100.0%	2/3	66.7%
	Female	5/55	9.1%	50/55	90.9%	38/55	69.1%
20-29	Male	2/24	8.3%	23/24	95.8%	9/24	37.5%
	Female	58/257	22.6%	232/257	90.3%	184/257	71.6%
30-39	Male	3/16	18.8%	12/16	75.0%	8/16	50.0%
	Female	57/189	30.2%	170/189	89.9%	145/189	76.7%
40-49	Male	6/11	54.5%	11/11	100.0%	8/11	72.7%
	Female	14/44	31.8%	39/44	88.6%	30/44	68.2%
50-59	Male	1/10	10.0%	9/10	90.0%	5/10	50.0%
	Female	3/11	27.3%	10/11	90.9%	10/11	90.9%
60-69	Male	3/6	50.0%	6/6	100.0%	4/6	66.7%
	Female	3/4	75.0%	4/4	100.0%	3/4	75.0%
70+	Male	0/0	0.0%	0/0	0.0%	0/0	0.0%
	Female	1/1	100.0%	1/1	100.0%	1/1	100.0%
UnkownAge	Male	0/0	0.0%	0/0	0.0%	0/0	0.0%
	Female	2/13	15.4%	9/13	69.2%	10/13	76.9%

Observed Prevalence in Individuals Undergoing ToRCH Antibody Assessment

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UnknownGender/Age	0/3	0.0%	1/3	33.3%	3/3	100.0%
Total	160/651	24.6%	582/651	89.4%	459	71.0%

Observed Prevalence in Pregnant Women

Prevalence of Analytes in Pregnant Women
	Toxoplasma		Rubella		CMV		HSV 1		HSV 2
Age	Pos/Total	% Prevalence	Pos/Total	% Prevalence	Pos/Total	% Prevalence	Pos/Total	% Prevalence	Pos/Total	% Prevalence
16-19	2/23	9.2%	23/23	100.0%	16/23	69.6%	15/23	65.2%	3/23	13.0%
20-29	12/131	27.0%	35/39	89.7%	104/131	79.4%	92/124	74.2%	51/124	41.1%
30-39	10/37	55.6%	127/129	98.4%	26/37	70.3%	30/36	83.3%	14/36	38.9%
40-49	5/9	8.3%	9/9	100.0%	5/9	55.6%	8/9	88.9%	7/9	77.8%
Total	29/200	14.50%	194/200	97.90%	151/200	75.50%	160/200	80.00%	75/200	37.50%

Observed Prevalence in Sexually Active Adults

			HSV 1	HSV 2
Age	Gender	Pos/Total	ర్యంPrevalence	Pos/Total	ళ్ళPrevalence
17-19	Male	. 0/0	0.0%	0/0	0.0%
	Female	27/38	71.1%	6/38	15.8%
20-29	Male	6/23	26.1%	1/23	4.3%
	Female	180/246	73.2%	57/245	23,3%
30-39	Male	10/16	62.5%	3/15	20.0%
	Female	165/187	88.2%	52/187	27.8%
40-49	Male	10/11	90.9%	3/11	27.3%
	Female	31/44	70.5%	16/44	36.4%
ર૦-રેવ	Male	5/10	50.0%	5/10	50.0%
	Female	8/11	72.7%	6/11	54.5%
୧୦-୧୨	Male	4/6	66.7%	2/6	33.3%
	Female	4/4	100.0%	2/4	50.0%
Total		450/596	75.5%	153/594	25.8%

HSV Hypothetical Predictive Values by Prevalence

Sexually Active Adults	Expectant Mothers
HSV-1	HSV-2	HSV-1	HSV-2

પપ

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Prevalence	PPV	NPV	PPV	NPV	PPV	NPV	PPV	NPV
50.0%	94.8%	98.5%	93.7%	96.8%	87.0%	99.2%	92.9%	97.0%
40.0%	92.4%	99.0%	90.9%	97.8%	81.7%	99.5%	89.7%	98.0%
30.0%	88.7%	99.4%	86.5%	98.6%	74.2%	99.6%	84.9%	98.7%
25.0%	85.9%	99.5%	83.2%	98.9%	69.1%	99.7%	81.4%	99.0%
20.0%	82.0%	99.6%	78.8%	99.2%	62.6%	99.8%	76.6%	99.2%
15.0%	76.3%	99.7%	72.5%	99.4%	54.2%	99.9%	69.8%	99.5%
10.0%	67.0%	99.8%	62.4%	99.6%	42.7%	99.9%	59.3%	99.7%
5.0%	49.0%	99.9%	44.0%	99.8%	26.1%	100.0%	40.8%	99.8%

Performance in Prospectively Collected Intended Use Populations

The performance of the ToRCH panel was evaluated using prospectively collected frozen remnant serum samples from a total of 651 individuals for which ToRCH IgG panel or testing for each of the individual analytes was ordered. Outside investigators tested 300 and 351 samples respectively.

Summary of Performance Characteristics In Individuals Undergoing ToRCH Antibody Assessment
		Predicate
		Positive	Equivocal	Negative	Site Total	PPA	95% CI
Toxoplasma	Positive	136	3	16	155	99.3% (136/137)	96.0% - 100%
	Equivocal	0	0	3	3
	Negative	0	1	490	491	95.7% (450/514)	98.0% - 99.9%
	Invalid	0	0	2	2
	Site Total	136	4	511	651
Rubella	Positive	533	4	4	541	98.5% (533/541)	97.1% - 99.4%
	Equivocal	3	1	1	5
	Negative	2	3	60	65	*87% (60/69)	76.7% - 93.9%
	Invalid	0	0	0	0
	Site Total	538	8	65	611
CMV	Positive	450	6	6	462	99.6% (450/452)	98.4% - 100%

of Performance Characteristics In Individuals Undergoing ToRCH Antibody Assessment summ

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Equivocal	1	2	4	7
Negative	1	0	181	182	91.3% (181/197)	87.2% - 95.3%
Invalid	0	0	0	0
Site Total	452	8	191	651

*4/4 discrepant Rubella samples which tested positive by ELSA had low positive values for AheMA and high negative values for ELISA, 4/4 discrepant Rubella samples which tested positive by AtheNA and equivocal by EUSA had low positive values for El.SA.

HSV1 and HSV2 Performance in Sexually Active Adults

The performance of the HSV1 and HSV2 was evaluated in prospectively collected samples using results from 596/651 individuals between the ages of 17 and 69.

		Positive	Equivocal	Negative	Site Total	SensitivitySpecificity	95% Cl
	HSV 1
	Positive	418	0	8	426	98.6% (418/424)	97.0% - 99.5%
	Equivocal	4	0	1	5
	Negative	2	0	163	165	94.6% (163/172)	90.3% - 97.6%
	Invalid	0	0	0	0
AthenA Multi-Lyte IgG Plus	Site Total	424	0	172	596
	HSV 2
	Positive	127	0	27	154	96.9% (127/131)	92.4% - 98.8%
	Equivocal	1	0	3	4
	Negative	3	0	433	436	93.5% (433/463)	90.9% - 95.6%
	Invalid	0	0	0	0
	Site Total	131	0	463	594*
Equivocal	0	0	1	1
Negative	0	0	170	170	95.3% (170/178)	91.3% - 98.0%
Invalid	0	0	0	0
Site Total	22	1	177	200
Rubella
Positive	194	0	0	194	99.0% (194/196)	96.4% - 99.9%
Indeterminate	1	0	0	1
Negative	0	1	4	5	100.0% (4/4)	47.3% - 100%
Invalid	0	0	0	0
Site Total	195	1	4	200
CMV
Positive	151	0	0	151	98.1% (151/154)	94.4% - 99.6%
Equivocal	0	0	0	0
Negative	0	3	46	49	100.0% (46/46)	93.7% - 100%
Invalid	0	0	0	0
Site Total	151	3	46	200
HSV 1
Positive	137	0	8	145	99.3% (137/138)	96.1% - 100%
Equivocal	0	0	0	0
Negative	1	0	46	47	85.2% (46/54)	72.3% - 93.4%
Invalid	0	0	0	0
Site Total	138	0	54	192
HSV 2
Positive	68	0	7	75	97.1% (68/70)	90.1% - 99.7%
Equivocal	0	0	2	2
Negative	2	0	113	115	92.6% (113/122	86.5% - 96.6%
Invalid	0	0	0	0
Site Total	70	0	122	192
	CDC Result
		Positive	Negative	Site Total	PPANPA	95% CI
	Toxoplasma
	Positive	70	0	70	100.0% (70/70)	95.8% - 100.0%
	Equivocal	0	0	0
	Negative	0	30	30	100.0% (30/30)	90.5% - 100.0%
	Invalid	0	0	0
	Site Total	70	30	100
	Rubella
	Positive	80	0	80	100.0% (80/80)	96.3% - 100.0%
	Equivocal	0	0	0
	Negative	0	20	20	100.0% (20/20)	86.1% - 100.0%
	Invalid	0	0	0
AtheNA Multi-Lyte TORCH IgG Plus	Site Total	80	20	100
	CMV
	Positive	52	2	54	100.0% (52/52)	94.4% - 100.0%
	Equivocal	0	0	0
	Negative	0	46	46	95.8% (46/48)	90.2% - 100.0%
	Invalid	0	0	0
	Site Total	52	48	100
	HSV 1
	Positive	50	0	50	100.0% (50/50)	94.2% - 100.0%
	Equivocal	0	0	0
	Negative	0	50	50	100.0% (50/50)	94.2% - 100.0%

Summary of Performance Characteristics in Sexually Active Adults

Performance in Pregnant Women Population

Zeus Scientific internally evaluated 200 frozen remnant serum samples collected from pregnant women between the ages of 15 and 46 for which ToRCH antibody testing was requested.

Image /page/9/Figure/8 description: This image is a table that summarizes the performance characteristics of a population of pregnant women. The table shows the results of a Toxoplasma test, with columns for positive, equivocal, and negative results. The table also includes columns for site total, NPA, and 95% CI. For the positive Toxoplasma test, the table shows 22 positive results, 1 equivocal result, and 6 negative results, with a site total of 29. The NPA is 100.0% (22/22), and the 95% CI is 87.3%-100%.

ની

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Agreement with CDC Panel

The performance of the AtheNA Multi-Lyte ToRCH IgG Plus test system was assessed using masked, well characterized serum panel from the CDC. The panels consist of:

1. 70% Toxo positive and 30% Toxo negative samples.
  14 47

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1. 80% Rubella positive and 20% Rubella negative samples.
1. 54% CMV positive and 46% CMV negative samples.
1. 24% HSV1 and HSV2 dual-positive samples, 50% HSV1 positive and 50% HSV1 negative samples and 48% HSV2 positive and 52% HSV2 negative samples.

The results are presented to convey further information on the performance of the test kit and do not imply endorsement of the assay by the CDC.

{12}------------------------------------------------

Invalid	0	0	0
Site Total	50	50	100
HSV 2
Positive	48	1	49	100.0% (48/48)	94.0% - 100.0%
Equivocal	0	0	0
Negative	0	51	51	98.1% (51/52)	94.3% - 100.0%
Invalid	0	0	0
Site Total	48	52	100

HSV 1 & 2 Performance in a Low Prevalence Population

The relative specificity of HSV 1 & 2 was assessed internally using sera from a low prevalence population. The low prevalence population was comprised of serum samples from 18 and 19 year old subjects previously tested for infections considered non-sexual in nature.

HSV-1 Reactivity: The predicate immunoblot device was positive for 7 samples and negative for 60 samples. The AtheNA Multi-Lyte HSV 1& 2 IgG test system agreed with 85.7% (6/7) of immunoblot positives and 98.3% (59/60) of immunoblot negatives.

HSV-2 Reactivity: The predicate immunoblot device was positive for 0 samples and negative for 67 samples. The AtheNA Multi-Lyte HSV 1& 2 IgG test system agreed with 100% (0/0) of immunoblot positives and 100% (67/67) of immunoblot negatives.

Performance in Low Prevalence Population
AtheNA Multi-Lyte IgG Plus				Predicate
		Positive	Equivocal	Negative	Site Total	PPANPA	95% CI
	HSV 1
	Positive	8	0	2	10	100.0% (8/8)	68.8% - 100%
	Equivocal	0	0	0	0
	Negative	0	0	56	56	96.6% (56/58)	88.1% - 99.6%
	Invalid	0	0	0	0
	Site Total	8	0	58	66
	HSV 2
	Positive	3	0	1	4	100.0% (3/3)	36.8% - 100%
	Equivocal	0	0	0	0
	Negative	0	0	62	62	98.4% (62/63)	91.5% - 100%
	Invalid	0	0	0	0

Performance in Low Prevalence Population

{13}------------------------------------------------

----------------------------------------------------------------------------	Site Total		63	ee
					------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

Rubella Retrospective Negative Sample Study

The relative specificity of Rubella was assessed internally at Zeus using pre-selected banked samples of sera which previously tested negative for Rubella antibody by the predicate device.

Rubella Reactivity: The predicate ELISA device was positive for 0 samples and negative for 100 samples. The Rubella analyte in the AtheNA Multi-Lyte ToRCH IgG Plus test system agreed with 100% (0/0) of ELISA positives and 100% (100/100) of ELISA negatives.

	Rubella Retrospective Negative Sample Study
	Predicate
		Positive	Equivocal	Negative	Site Total	PPANPA	95% CI
AtheNA Multi-Lyte IgG Plus	Rubella
	Positive	0	0	0	0	N/A	N/A
	Indeterminate	0	0	0	0
	Negative	0	0	100	100	100.0% 100/100)	97.1% - 100%
	Invalid	0	0	0	0
	Site Total	0	0	100	100

Clinical performance with International Standards

Performance of the AtheNA Multi-Lyte ToRCH IgG Plus was assessed using the CDC Reference Panels. Traceability and linearity was demonstrated for Rubella using the WHO standard. The performance of Rubella IgG was demonstrated using the CDC low titer standard.

Rubella Traceability to WHO Standard

Traceability was investigated internally to assess the device's correlation to the WHO Standard at the cut-off. The Standard was diluted and tested in duplicate with the device on two lot numbers and the Percent Recovery calculated. At the WHO Standard dilution of 10.63, the mean of the results on Lot 1 was 9.1 with a recovery of 96%. The mean of results for Lot 2 was 10 with a recovery of 102%.

Expected	AtheNA Multi-Lyte lot 1			AtheNA Multi-Lyte lot 2
ResultIU/mL	Measured Mean		Mean % Recovery		Measured Mean		Mean % Recovery
	Result	IU/mL	Result	IU/mL	Result	IU/mL	Result	IU/mL
1.33	3	2.7	210	205	3	2.7	231	205
2.66	4	3.6	148	137	4	3.6	146	137
5.31	6	5.5	110	103	6	5.5	105	103

{14}------------------------------------------------

10.63	10	9.1	કેટ	સેસ્ટ		10	102	ઉત્ત
21.25	20	18.2		કેટ	20	18.2	92	કેટ
----------------------------------------------------------------------------------------------------------------------	14-44-4-4.					-------	I Am and Call Compression and

Rubella Low Range Study

A Rubella IgG low range linearity study was performed to demonstrate linearity across the lower range of the assay using the WHO Standard was diluted in duplicate, the means established and the percent recovery of the expected results calculated.

Image /page/14/Figure/3 description: The image is a graph titled "Rubella Low Range Linearity Study". The graph plots "Observed Result, U/mL" on the y-axis and "Predicted Result, IU/mL" on the x-axis. A linear trendline is plotted on the graph, and the equation for the trendline is y = 0.8774x + 1.3999, with an R-squared value of 0.9977.

Rubella Performance with CDC Low Titer Standard

Rubella performance was assessed with the CDC low titer sample (21 IU/mL). The sample was aliquoted, diluted in duplicate and tested by three technicians. Percent recovery was calculated for both neat and diluted samples

CDC Low Titer Rubella Standard: 21 IU/mL
Tech	Neat	Interpretation	% Recovery	1:2 Dilution	Interpretation	% Recovery
1	22	Positive	102%	11	Positive	102%
1	20	Positive	93%	10	Positive	99%
2	22	Positive	103%	11	Positive	109%
2	22	Positive	105%	11	Positive	104%
3	20	Positive	95%	11	Positive	101%
3	21	Positive	100%	12	Positive	111%

Rubella Performance With CDC Low Titer Sample

Precision and Reproducibility

Assay precision and reproducibility was evaluated internally and at two external clinical sites. The study was conducted as follows: A panel of six samples was identified and/or prepared (by Zeus Scientific, Inc.) for use in the study based upon their activity on the AtheNA Multi-Lyte assay. Two samples of the panel

18 57

{15}------------------------------------------------

were negative, two were high positives and the other two samples were near the assay cut off for each specific target. To assess reproducibility, each sample was aliquoted twice and each aliquot was run in triplicate, each day. This results per day. This was repeated for three days at each site and the resulting data used to assess precision at each facility

	Sample	Mean	Within-Run		Between-Day		Between-Run		Between-Site		Total
Member	N	AU/mL	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Toxo IgG Positive 1	54	747.2	52.1	7.3	61.6	8.5	34.5	4.7	71.5	8.7	157.2	8.0
Toxo IgG Positive 2	54	770.3	50.8	6.8	63.1	8.3	39.2	4.9	78.7	8.5	170.3	8.6
Toxo IgG Positive 1 (near Cut-off)	54	158.8	158.8	9.3	18.2	11.3	12.6	8.0	18.8	12.1	29.5	11.9
Toxo IgG Positive 2 (near Cut-off)	54	140.2	140.2	8.4	16.8	11.3	13.0	8.3	20.2	12.1	33.2	12.9
Toxo IgG Negative 1	54	10.5	10.5	40.9	3.8	39.7	1.3	15.0	4.3	38.4	5.0	40.7
Toxo IgG Negative 2	54	9.1	9.1	45.4	4.1	47.4	2.4	22.6	4.7	48.0	5.6	49.8
Rubella IgG Positive 1	54	1296.9	50.1	3.9	68.4	5.0	54.5	3.7	95.4	5.3	301.3	5.7
Rubella IgG Positivetive 2	54	1102.6	58.2	5.4	63.6	5.8	37.3	3.4	99.5	6.5	173.1	7.0
Rubella IgG Positive 1 (near Cut-off)	54	242.8	13.3	5.4	15.6	6.5	10.2	4.4	25.6	6.2	51.9	6.4
Rubella IgG Positive 2 (near Cut-off)	54	189.9	10.1	5.2	15.5	9.5	11.5	5.7	28.3	10.1	37.9	10.4
Rubella IgG Negative 1	54	27.9	4.8	20.5	16.1	21.9	1.8	8.3	4.2	21.3	19.4	19.3
Rubella IgG Negative 2	54	47.4	4.0	10.3	6.5	9.8	3.7	9.4	8.8	10.9	23.8	11.0
CMV IgG Positive 1	54	996.5	88.2	8.5	99.2	9.7	58.5	5.9	108.2	10.1	167.2	10.0
CMV IgG Positive 2	54	756.9	54.5	7.0	66.7	8.7	46.8	6.3	73.3	8.8	163	8.7
CMV IgG Positive 1 (near Cut-off)	54	119.2	10.1	8.2	13.2	11.1	9.6	8.3	14.9	10.9	18.3	10.1
CMV IgG Positive 2 (near Cut-off)	54	135.3	13.6	9.9	16.4	12.1	12.6	9.4	19.5	10.7	21.2	10.8
CMV IgG Negative 1	54	17.9	5.6	29.8	6.1	32.7	3.4	19.5	6.3	28.2	7.4	27.1
CMV IgG Negative 2	54	16.4	5.5	35.5	5.9	37.1	3.6	23.4	6.3	35.4	7.3	35.4
HSV 1 IgG Positive 1	54	310.1	24.2	7.9	24.8	8.1	10	3.3	31	8.9	32.3	10.1
HSV 1 IgG Positive 2	54	392.7	31.5	8.0	32.3	8.2	15.6	3.8	48.1	8.7	50.8	8.4
HSV 1 IgG Positive 1 (near Cut-off)	54	144.6	15.4	10.7	17.2	12.0	8.9	6.0	22.3	12.4	23.0	12.4
HSV 1 IgG Positive 2 (near Cut-off)	54	191.7	17.6	9.1	19.7	10.2	9.6	5.1	24.5	9.9	27.4	10.6
HSV 1 IgG Negative 1	54	26.4	3.9	15.5	3.9	15.5	1.4	5.2	4.8	16.2	6.1	16.8
HSV 1 IgG Negative 2	54	8.2	2.3	30.7	2.5	34.1	1.3	17.0	2.7	36.9	3.5	39.4
HSV 2 IgG Positive 1	54	445.7	26.9	6.1	40.1	8.9	33.8	7.4	53.4	9.2	58.1	9.1
HSV 2 IgG Positive 2	54	355.6	27.0	7.4	30.5	8.4	18.8	5.2	51.5	8	57.2	8.6
HSV 2 IgG Positive 1 (near Cut-off)	54	152.2	14.9	9.9	16.0	10.6	7.6	5.2	25.3	10.0	31.6	11.2
HSV 2 IgG Positive 2 (near Cut-off)	54	114.1	11.4	9.8	12.5	10.9	6.3	5.7	15.4	10.8	20.4	10.8
HSV 2 IgG Negative 1	54	16.9	4.0	29.7	4.3	31.3	2.0	12.1	4.3	35.8	7.0	41.0
HSV 1 IgG Negative 2	54	21.2	5.9	27.5	6.5	30.6	3.1	15.2	6.8	26.6	8.9	27.6

{16}------------------------------------------------

క్రశ 20

{17}------------------------------------------------

lot 09046891 Linearity Study

WHO Value	Rubella IgG			Rubella IgG			Mean(U/mL)	converted IU/mL
	AU	U/mL	converted IU/mL	AU	U/mL	converted IU/mL
21.25	198	18	17	245	22	20	20	18
10.63	122	11	10	101	9	8	10	9
5.31	57	5	5	71	7	6	6	5
2.66	45	4	4	41	4	3	4	4
1.33	25	2	2	36	3	3	3	3

WHO Value IU	Means U/mL
21.25	20
10.63	10
5.31	6
2.66	4
1.33	3

WHO Value IU	Converted Means IU/mL
21.25	18
10.63	9
5.31	5
2.66	4
1.33	3

Image /page/17/Figure/5 description: The image is a graph titled "Rubella IgG Low Range Linearity Study Rubella WHO Standard". The graph plots "Observed Result, U/mL" on the y-axis and "Predicted Result, IU/mL" on the x-axis. A linear trendline is plotted on the graph, with the equation y = 0.8774x + 1.3999 and R^2 = 0.9977.

{18}------------------------------------------------

Image /page/18/Picture/1 description: The image shows the seal of the Department of Health & Human Services - USA. The seal is circular, with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. In the center of the seal is an abstract image of an eagle.

Food and Drug Administration 10903 New Hampshire Avenue Document Mail Center - WO66-0609 Silver Spring, MD 20993-0002

JUL 1 6 2010

Zeus Scientific Inc. c/o Ms. Ewa Nadolczak Manager Clinical Affairs 200 Evans Way Branchburg, NJ 08876

Re: K093784

Trade/Device Name: AtheNA Multi-Lyte® ToRCH IgG Plus Test System Regulation Number: 21 CFR 866.3510 Regulation Name: Rubella Virus Serological Reagents. Regulatory Class: Class II Product Code: OPM, LGD, LFZ, MXJ, MYF Dated: July 12, 2010 Received: July 13, 2010

Dear Ms. Nadolczak:

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.

If your device is classified (see above) into class II (Special Controls), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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

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

This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5450. 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 Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely vours.

Sauza Hopkins

Sally A. Hoivat. M.S. Director Division of Microbiology Devices Office of In Vitro Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number:

2093784

Device Name: AtheNA Multi-Lyte® ToRCH IgG Plus Test System

Indications for Use:

The test is not intended for use in screening blood or plasma donors. The performance of this assay has not been established for use in a pediatric population, neonatal screening, immunocompromised or immunosuppressed patients or for use at point of care facilities.

Prescription Use X (Part 21 CFR 801 Subpart D)

AND/OR

Over-The-Counter Use (21 CFR 807 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)

the Schif

Division Sign-Of

Office of In Vitro Diagnostic Device Evaluation and Safety

Page 1 of

510(k) k 093784

Regulation Number and Section

§ 866.3510 Rubella virus serological reagents.

(a)
Identification. Rubella virus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to rubella virus in serum. The identification aids in the diagnosis of rubella (German measles) or confirmation of a person's immune status from past infections or immunizations and provides epidemiological information on German measles. Newborns infected in the uterus with rubella virus may be born with multiple congenital defects (rubella syndrome).(b)
Classification. Class II. The special controls for this device are:(1) National Committee for Clinical Laboratory Standards':
(i) 1/LA6 “Detection and Quantitation of Rubella IgG Antibody: Evaluation and Performance Criteria for Multiple Component Test Products, Speciment Handling, and Use of the Test Products in the Clinical Laboratory, October 1997,”
(ii) 1/LA18 “Specifications for Immunological Testing for Infectious Diseases, December 1994,”
(iii) D13 “Agglutination Characteristics, Methodology, Limitations, and Clinical Validation, October 1993,”
(iv) EP5 “Evaluation of Precision Performance of Clinical Chemistry Devices, February 1999,” and
(v) EP10 “Preliminary Evaluation of the Linearity of Quantitive Clinical Laboratory Methods, May 1998,”
(2) Centers for Disease Control's:
(i) Low Titer Rubella Standard,
(ii) Reference Panel of Well Characterized Rubella Sera, and
(3) World Health Organization's International Rubella Standard.