ZEUS SCIENTIFIC, INC. TREPONEMA PALLIDUM IGG TEST SYSTEM

K102283 · Zeus Scientific, Inc. · LIP · Feb 10, 2011 · Microbiology

Device Facts

• Record ID: K102283
• Device Name: ZEUS SCIENTIFIC, INC. TREPONEMA PALLIDUM IGG TEST SYSTEM
• Applicant: Zeus Scientific, Inc.
• Product Code: LIP · Microbiology
• Decision Date: Feb 10, 2011
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3830
• Device Class: Class 2

Intended Use

The ZEUS ELISA Treponema pallidum IgG Test System is intended for the qualitative detection of specific IgG class antibodies to T. pallidum in human serum. The test may be used in conjunction with non treponemal testing and clinical findings to provide serological evidence of infection with T. pallidum. This test is for in vitro diagnostic use only. This test is not intended for screening blood or plasma donors.

Device Story

ELISA-based in vitro diagnostic test for qualitative detection of IgG antibodies to Treponema pallidum in human serum. Principle: microwell strips sensitized with recombinant T. pallidum antigen; patient serum incubated; antigen-specific IgG binds to immobilized antigen; peroxidase-conjugated goat anti-human IgG added; substrate (TMB) added; color change measured photometrically. Performed in clinical laboratories by trained personnel. Output: optical density (OD) value converted to index value; results interpreted as negative (≤0.90), equivocal (0.91-1.1), or positive (>1.1). Aids clinicians in diagnosing syphilis infection when used alongside non-treponemal tests and clinical evaluation. Benefits: provides serological evidence of infection to guide clinical management.

Clinical Evidence

Clinical performance evaluated across 2,937 samples including prospective cohorts (500 patients with syphilis test ordered, 500 pregnant women, 1000 hospitalized patients) and retrospective banked samples (HIV-positive, RPR/TPPA positive, pregnant women, CDC syphilis panel). Compared against TPPA reference assay. Overall agreement varied by population; e.g., 99.2% NPA in prospective patients, 100% NPA in pregnant women. CDC panel agreement was 93.0% (146/157). Precision and reproducibility studies showed acceptable CVs (<15% for reactive, <25-50% for negative samples).

Technological Characteristics

ELISA immunoassay; polystyrene 96-well plate; recombinant p17 T. pallidum antigen; goat anti-human IgG (Fc chain specific) HRP conjugate; TMB substrate; photometric detection. Single-point calibration curve. Manual or automated processing. No specific materials standards cited.

Indications for Use

Indicated for qualitative detection of IgG antibodies to Treponema pallidum in human serum as an aid in syphilis diagnosis. Used with non-treponemal tests and clinical findings. Prescription use only. Not for blood/plasma donor screening.

Regulatory Classification

Identification

Treponema pallidum treponemal test reagents are devices that consist of the antigens, antisera and all control reagents (standardized reagents with which test results are compared) which are derived from treponemal sources and that are used in the fluorescent treponemal antibody absorption test (FTA-ABS), the Treponema pallidum immobilization test (T.P.I.), and other treponemal tests used to identify antibodies to Treponema pallidum directly from infecting treponemal organisms in serum. The identification aids in the diagnosis of syphilis caused by bacteria belonging to the genus Treponema and provides epidemiological information on syphilis.

Predicate Devices

• Pheonix Bio-Tech Trep-Chek

Submission Summary (Full Text)

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# Zeus Scientific Inc. 510(k) Summary ZEUS ELISA Treponema pallidum IgG FEB 1 0 2011 Test System K102283

Zeus Scientific, Inc. (Zeus) PO Box 38, Raritan, NJ 08869 [phone] Contact: Ewa Nadolczak, Manager of Clinical Affairs, Direct [phone] [e-mail]

Measurand: Treponema pallidum IgG antibodies. Type of Test: ELISA. Proprietary Name: ZEUS ELISA Treponema pallidum IgG Test System.

### Section 1: Regulatory Information

- Device Classification: Enzyme Linked Immunoabsorbent Assay, Treponema pallidum 1.
- 2. Regulation Description: Treponema pallidum treponemal test reagents
- 3. Class: 2
- 4. Product Code: LIP
- 5. Panel: Microbiology
- Regulation Number: 866.3830 ହ.

### Section 2: Intended Use

The ZEUS EUSA Treponema pallidum IgG Test System is intended for the qualitative detection of specific IgG class antibodies to T. pollidum in human serum. The test may be used in conjunction with non treponemal testing and clinical findings to provide serological evidence of infection with T. pallidum. This test is for in vitro diagnostic use only.

This test is not intended for screening blood or plasma donors.

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### Section 3: Indications for Use

Indications for Use: This test system when used in conjunction with non-treponemal based assays provides serological evidence of infection with Treponema pallidum.

Special Conditions for Use Statements:

This test is for in vitro diagnostic use only. · This test is for prescription use only. This test is not intended for screening blood or plasma donors.

### Section 4: Substantial Equivalence

Examination of enclosed data indicates that the ZEUS ELISA Treponema pallidum IgG Test System for the qualitative detection of specific human IgG class antibodies to treponema pallidum in human serum is substantially equivalent to a commercially marketed test system which has been previously cleared by the FDA for in vitro diagnostic use.

| T. pallidum IgG Result Interpretation: | | |
|----------------------------------------|-----------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| OD Value | Result | Interpretation |
| ≤ 0.90 | Negative | A result of ≤0.90 indicates no detectable IgG antibodies to T. pallidum and should be reported<br>as non-reactive for IgG antibody to T. pallidum. |
| 0.91 - 1.1 | Equivocal | Specimens with results in the equivocal range (0.91 - 1.1) should be retested in duplicate. Any<br>two of the three results which agree should be reported. Specimens that remain equivocal<br>after repeat testing may be tested by an alternate serologic procedure and/or re-evaluated by<br>drawing another sample one to three weeks later. |
| >1.1 | Positive | A result of > 1.1 indicates that the specimen is positive for IgG antibody to T. pallidum, the<br>causative agent for syphilis. |
| | | A positive test result presumes a current or past infection with T. pallidum, and should be<br>reported as reactive for IgG antibody to T. pallidum. Other T. pallidum serology assays should<br>be performed to confirm or rule out a current case of active syphilis. |

### Section 5: Interpretation of Results

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## Section 5A: Comparison of Investigational Device to Predicate Device

The comparisons of the ZEUS ELISA T.pallidum IgG Test System to the predicate device follow, including intended use and various aspects of the procedure.

| Characteristic | ZEUS ELISA T.pallidum IgG | Pheonix Bio-Tech Trep-Chek |
|----------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Use<br>Intended Use | For in vitro diagnostic use only<br>Intended Use: The ZEUS ELISA Treponema<br>pallidum ELISA Test System is intended for<br>the qualitative detection of specific IgG class<br>antibodies to T. pallidum in human serum.<br>The test system is intended to be used as an<br>aid in the laboratory diagnosis of Syphilis<br>disease caused by T. pallidum spirochete. | For in vitro diagnostic use only<br>The Phoenix Bio-Tech Corp. Syphilis Trep-<br>Chek Test Kit is a confirmatory immunoassay<br>for the qualitative detection of Treponema<br>pallidum IgG antibodies in human serum. |
| Assay<br>Detection Method<br>Solid Phase<br>Antigen Used | Immunoassay<br>Colormetric<br>Polystyrene 96 well plate<br>recombinant p 17 Treponema pallidum<br>antigen | Immunoassay<br>Colormetric<br>Polystyrene 96 well plate<br>Recombinant Treponema pallidum antigen |
| Specimen Tested | Human Serum | Human Serum |
| Controls | One Positive Control, one Negative Control | One Positive Control, one Negative Control |
| Calibration | Includes Calibrator (human serum) | Includes Calibrator (human serum) |
| Analyte Measured<br>Sample Dilution | Human IgG<br>1:21 in SAVe Diluent | Human IgG<br>1:20 in phosphate buffer based diluent |
| Sample Incubation | 25 +/- 5 minutes at room temperature | 30 +/- 2 minutes at room temperature |
| Post Sample Wash | 5x wash (dispense/aspirate) | 4x wash (dispense/aspirate) |
| Conjugate | Goat anti-human IgG; Fc chain specific | Goat anti-human IgG; y chain specific |
| Conjugate Label | Horse radish peroxidase | Horse radish peroxidase |
| Conjugate Incubation | 25 +/- 5 minutes at room temperature | 30 +/- 2 minutes at room temperature |
| Post Conjugate Wash | 5x wash (dispense/aspirate) | 4x wash (dispense/aspirate) |
| Substrate | TMB | TMB |
| Reading<br>Data Points | Read the color change (optical density) of<br>the wells<br>Read one OD value for each control and<br>sample | Read the color change (optical density) of<br>the wells<br>Read one OD value for each control and<br>sample |
| Math | Single point curve | Single point curve |
| Scale | Calculate the index value of unknown<br>samples by comparing their OD to the cut<br>off OD | Calculate the index value of unknown<br>samples by comparing their OD to the cut<br>off OD |
| Interpretation Criteria | Negative is <= 0.90, Positive is >= 1.10 and<br>Equivocal is 0.90 - 1.09 | Negative is <= 0.90, Positive is >= 1.10 and<br>Equivocal is 0.90 - 1.09 |

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### Section 6: Test Principle

The ZEUS ELISA Treponema pallidum IgG Test System is designed to detect IgG class antibodies in human sera to Treponema pallidum. Wells of plastic microwell strips are sensitized by passive absorption with T. pallidum antigen. The test procedure involves three incubation steps:

- 1. Test sera are diluted with SAVE Diluent. During sample incubation, any antigen specific lgG antibody in the sample will bind to the immobilized antigen. The plate is washed to remove unbound antibody and other serum components.
- 2. Peroxidase conjugated goat anti-human IgG is added to the wells and the plate is incubated. The conjugate will react with IgG antibody immobilized on the solid phase in step 1. The wells are washed to remove unbound conjugate.
- 3. The microwells containing immobilized peroxidase conjugate with peroxidase substrate solution. Hydrolysis of the substrate by peroxidase produces a color change. After ten minutes have elapsed, the reaction is stopped and the color intensity of the solution is measured photometrically. The color intensity of the solution depends on upon the antibody concentration in the original test sample.

### Section 7: Linearity

The WHO International Standard was diluted serially. Each dilution was tested in duplicate, the mean calculated and the result plotted.

Image /page/3/Figure/7 description: The image is a graph titled "T.pallidum IgG Linearity". The x-axis is labeled "Numerical Interpretation of Dilution Factor" and ranges from 0 to 0.6. The y-axis is labeled "Mean Index Value" and ranges from 0.00 to 3.50. The graph shows a linear relationship with an R-squared value of 0.9971.

### Section 8: Analytical Specificity

### Interfering Substances

The effect of potential interfering substances on sample results generated using the test system was evaluated with the following possible interfering substances: albumin, bilirubin, cholesterol, hemoglobin, triglycerides and intralipids.

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The quantity of analyte in each interfering substance is as follows:

Bilirubin: 1mg/dL (low), 15 mg/dL (high) 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 for Treponema pallidum IgG were chosen based on their performance on the Treponema pallidum IgG ELISA test system: positive, borderline and negative. The samples were exposed to the possible interfering substances and tested. An increase or reduction of signal equal to or less than 20% is considered acceptable. The negative sample may have a signal change greater than 20% if there is no change in the qualitative result of the sample. All signal changes greater than 20 % must be specified in the package insert.

All positive samples showed less than a 20% recovery of signal.

The borderline samples showed a recovery of signal <20% of with the exception of the high spike of hemoglobin (25.2%).

The negative sample showed a change of signal (>20%) with the high and low spikes of albumin, hemoglobin, intralipid, bilirubin, cholesterol and triglycerides. The negative sample results in each instance stayed below the cut-off and the change in signal did not affect the qualitative result.

#### Cross Reactivity

Ten samples minimally for each cross-reactant were tested. The results presented were obtained by testing the analytes against high concentrations of possible cross reactants. The results of this study are summarized in the following table.

| Treponema pallidum IgG<br>Cross reactivity Study | |
|--------------------------------------------------|-----------------|
| Analyte | positive/tested |
| EBV | 0/10 |
| ANA | 0/10 |
| RF IgM | 0/10 |
| Rubella | 0/10 |
| HIV | 0/10 |
| HSV 1 | 0/10 |
| HSV 2 | 0/10 |
| Sera from Pregnant Patients | 0/10 |
| Hepatitis B | 0/10 |
| VZV | 0/10 |
| VZV IgM | 0/10 |
| CMV | 0/10 |
| Toxoplasma | 0/10 |
| Lyme G/M | 0/10 |
| Hepatitis C | 0/10 |

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#### Detection Limits

This is a qualitative assay (Positive, Negative or Equivocal results) so there is no limit of detection study.

### Section 9: Clinical Performance

#### Clinical Data Generated for Submission: Method Comparison with Predicate Device

- 1. Prospectively Collected Intended Use Populations:
- . 500 unselected samples from patients with a syphilis test ordered. The samples were submitted for syphilis antibody testing, sequentially numbered, de-identified and archived.
- . 500 unselected samples from pregnant women with a syphilis test ordered. The samples were submitted for syphilis antibody testing, sequentially numbered, de-identified and archived.

After procurement, the samples were tested at a hospital laboratory located in the Mid-Atlantic, the Northeast and at Zeus. The hospital laboratories tested 200 samples from patients with a syphilis test ordered and 200 samples from pregnant women. Zeus tested 100 samples from patients with a syphilis tests ordered and 100 samples from pregnant women.

- 2. Prospectively Collected Population of Unselected Hospitalized Patients: Additional clinical performance was assessed in a population of 1000 hospitalized patients. These samples were pulled from a hospital laboratory routine workload of patient testing.
- 3. Prospective HIV-1 Positive Samples: 223 banked known positive HIV-1 samples were acquired from a New York vendor.
- Retrospective TPPA /RPR Positive: 280 samples requested to be RPR/TPPA positive were 4. purchased from a New York vendor.
- 5. Retrospective TPPA Negative Collected from Pregnant Women: 250 samples requested to be collected from pregnant women and requested to be syphilis antibody negative were purchased from a New York vendor.
- Retrospective TPPA Positive Collected from Pregnant Women: 27 samples requested to be 6. collected from pregnant women and requested to be RPR/TPPA positive were purchased from a New York vendor.
- 7. CDC Syphilis Panel: 157 samples of various reactivity to syphilis were evaluated.

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- 8. Precision and Reproducibility: Precision of the device was assessed using fifteen samples. These repeatability studies were performed internally at the manufacturing site's laboratory. Reproducibility was assessed using fifteen samples tested at two external sites and internally at Zeus.
#### Prevalence and Expected Values

To determine expected values in the populations tested, internal investigators assessed the device's performance with 500 masked samples prospectively collected from individuals and 500 samples from pregnant women. The samples were requested to be random, unselected sera submitted for syphilis antibody testing. Additional studies were conducted in a population of 1000 unselected hospitalized patients.

In the 500 prospectively collected samples from patients ranging in age from <1 to >70 years of age, 7 tested positive. The overall observed prevalence in this group was 1.4% (7/500 samples).

In the 500 samples collected from pregnant women ranging in age from 15 to 48, 3/498 samples tested positive. The observed prevalence in this group was 0.6%. Two samples were excluded due to questionable age.

In the group of 1000 samples from unselected hospitalized patients ranging in age from <1 to >70 years of age, 32 tested positive. The overall observed prevalence in this group was 3.2% (32/999 samples). One sample was QNS for testing.

#### Clinical Performance and Comparative Testing for Intended Use Populations in a Prospective Study

The comparative study for the Intended Use Populations consisted of 500 unselected serum samples from patients with a syphilis test ordered. 500 purchased serum samples from pregnant women were also tested.

#### PERFORMANCE CHARACTERISTICS

The clinical study consisted of 2,937 serum samples evaluated at three sites located in the United States. All serum samples evaluated for concordance were tested with the treponemal (TPPA) reference assay. Samples that were positive by TPPA were reference assay positive. Samples that were negative by TPPA were reference assay negative. The testing sites, sample volumes are presented in the following table:

| Sites, Populations and Amounts of Samples Tested | | | | | | | | | |
|--------------------------------------------------|---------------------|---------------|--------------|--------------|----------------------|----------------|----------------|---------------|--|
| | Prospective Samples | | | | Rerospective Samples | | | | |
| | patients with | pregnant | sera from | purchased | purchased | purchased | sera requested | CDC | |
| | syphilis test | women with | unselected | sera from | sera from | sera requested | to be | Syphilis | |
| | test | syphilis test | hospitalized | HIV positive | pregnant | to be | | characterized | |

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| Sites | ordered | ordered | patients | patients | women | RPR/TPPA + | samples |
|----------|---------|---------|----------|----------|-------|------------|---------|
| Zeus | 100 | 100 | 350 | 223 | 0 | 0 | 0 |
| Monmouth | 200 | 200 | 350 | 0 | 277 | 280 | 0 |
| Mainline | 200 | 200 | 300 | 0 | 0 | 0 | 0 |
| CDC | 0 | 0 | 0 | 0 | 0 | 0 | 157 |
| Total | 500 | 500 | 1000 | 223 | 277 | 280 | 157 |

#### Prospectively Collected Intended Use Populations:

500 unselected samples from patients with a syphilis test ordered. The samples were submitted for syphilis antibody testing, sequentially numbered, de-identified and archived.

| | | Banked sera from patients with syphilis test ordered | | | | | |
|----------------------|------------|------------------------------------------------------|-----------|----------|------------|---------|------------|
| | | Predicate | | | | | |
| | | Positive | Equivocal | Negative | Site Total | PPA/NPA | 95% CI |
| T.pallidum IgG ELISA | Positive | 4 | 0 | 3 | 7 | 80.0% | 28.4-99.5% |
| | Equivocal | 1 | 0 | 1 | 2 | | |
| | Negative | 0 | 0 | 491 | 491 | 99.2% | 97.9-99.8% |
| | Site Total | 5 | 0 | 495 | 500 | | |

500 unselected samples from pregnant women with a syphilis test ordered. The samples were submitted for syphilis antibody testing, sequentially numbered, de-identified and archived.

| Banked purchased sera from pregnant women with syphilis test ordered | | | | | | | |
|----------------------------------------------------------------------|------------|----------|-----------|----------|------------|------------|------------|
| Predicate | | | | | | | |
| | | Positive | Equivocal | Negative | Site Total | PPA<br>NPA | 95% CI |
| T.pallidum IgG ELISA | Positive | 3 | 0 | 0 | 3 | 75.0% | 19.4-99.4% |
| | Equivocal | 0 | 0 | 0 | 0 | | |
| | Negative | 1 | 0 | 494 | 495 | 100.0% | 99.4-100% |
| | Site Total | 4 | 0 | 494 | 498 | | |

*2 samples excluded due to unverifiable age

Prospectively Collected Population of Unselected Hospitalized Patients: Additional clinical performance was assessed in a population of 1000 hospitalized patients. These samples were pulled from a hospital laboratory routine workload of patient testing and were tested at the three clinical sites.

Image /page/7/Figure/8 description: The image shows a table with three rows. The first row contains the text "Unselected hospitalized patients". The second row contains the text "Predicate". The third row contains the text "PPA".

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| | | Positive | Equivocal | Negative | Site Total | NPA | 95% CI |
|----------------------|-----------|----------|-----------|----------|------------|-------|------------|
| T.pallidum IgG ELISA | Positive | 13 | 1 | 18 | 32 | 61.9% | 38.4-81.9% |
| | Equivocal | 1 | 0 | 9 | 10 | | |
| | Negative | 7 | 0 | 950 | 957 | 97.1% | 95.9-98.1% |
| | QNS | 0 | 0 | 1 | 1 | | |
| Site Total | | 21 | 1 | 978 | 1000 | | |

Retrospective HIV-1 Positive Samples: 223 banked known positive HIV-1 samples were acquired from a New York vendor.

| | | Banked purchased known HIV-1 positive serum samples | | | | | |
|-------------------------|------------|-----------------------------------------------------|-----------|----------|------------|------------|------------|
| | | Predicate | | | | | |
| | | Positive | Equivocal | Negative | Site Total | PPA<br>NPA | 95% CI |
| T.pallidum IgG<br>ELISA | Positive | 41 | 0 | 1 | 42 | 85.4% | 72.2-93.9% |
| | Equivocal | 1 | 0 | 0 | 1 | | |
| | Negative | 4 | 2 | 174 | 180 | 99.4% | 96.9-100% |
| | Site Total | 46 | 2 | 175 | 223 | | |

Retrospective TPPA /RPR Positive: 280 samples requested to be RPR/TPPA positive were purchased from a New York vendor.

| | Banked purchased sera requested to be RPR/TPPA reactive | | | | | | |
|-------------------------|---------------------------------------------------------|-----------|----------|------------|------------|--------|------------|
| | Predicate | | | | | | |
| | Positive | Equivocal | Negative | Site Total | PPA<br>NPA | 95% CI | |
| T.pallidum IgG<br>ELISA | Positive | 259 | 1 | 4 | 264 | 98.5% | 96.2-99.6% |
| | Equivocal | 1 | | | 1 | | |
| | Negative | 3 | | 12 | 15 | 70.6% | 46.9-98.7% |
| | Site Total | 263 | 1 | 16 | 280 | | |

Retrospective TPPA Negative and TPPA Positive Samples Collected from Pregnant Women: 250 samples requested to be collected from pregnant women and requested to be syphilis antibody negative were purchased from a New York vendor. Only 27 samples requested to be collected from pregnant women and requested to be RPR/TPPA positive were available. These samples were purchased from a New York vendor.

> Banked purchased sera from pregnant women requested to be TPPA positive (27)

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| | | RPR/TPPA non-reactive (250) | | | | | |
|----------------------|------------|-----------------------------|-----------|----------|------------|---------|------------|
| | | Predicate | | | | | |
| | | Positive | Equivocal | Negative | Site Total | PPA/NPA | 95% CI |
| T.pallidum IgG ELISA | Positive | 26 | 1 | 0 | 27 | 92.9% | 76.5-99.1% |
| | Equivocal | 0 | 0 | 0 | 0 | | |
| | Negative | 2 | 0 | 248 | 250 | 99.6% | 97.8-100% |
| | Site Total | 28 | 1 | 248 | 277 | | |

CDC Syphilis Panel: 157 samples from patients with various clinical diagnoses with syphills disease were evaluated. The % agreement of the Treponema pallidum IgG ELISA kit with the clinical diagnosis is presented in the following table.

| Clinical Diagnosis | T. pallidum IgG ELISA Results | | | | % Agreement with Clinical Diagnosis<br>Presented with 95% CI | |
|---------------------|-------------------------------|-----------|----------|-------|--------------------------------------------------------------|------------|
| | Positive | Equivocal | Negative | Total | | |
| Primary Treated | 11 | 0 | 0 | 11 | 100% (11/11) | 76.2-100% |
| Secondary Untreated | 41 | 0 | 2 | 43 | 95.3% (41/43) | 84.2-99.4% |
| Secondary Treated | 39 | 0 | 0 | 39 | 100% (39/39) | 92.6-100% |
| Latent Untreated | 6 | 0 | 5 | 11 | 54.5% (6/11) | 23.4-83.3% |
| Latent Treated | 48 | 0 | 2 | 50 | 96.0% (48/50) | 86.3-99.5% |
| Congenital | 1 | 1 | 1 | 3 | 33.3% (1/3) | 0.84-90.6% |
| Total | 146 | 1 | 10 | 157 | 93.0% (146/157) | 87.8-96.5% |

#### PRECISION and REPRODUCIBILITY

#### Precision

Precision was evaluated at the manufacturer site. The study was conducted as follows: fifteen samples were identified and/or prepared (by Zeus Scientific, Inc.) for use in the study based upon their activity on the ZEUS ELISA Treponema pallidum IgG assay. Three samples each were selected that were negative, high negative, near cut-off, low positive. On each day of testing, the samples were diluted twice and tested. This was repeated in a second run on the same day by a different technologist for a total of twelve days. Precision is considered acceptable for the reactive samples if the Total CV is <15%. The reproducibility for the negative samples is considered acceptable if the Total CV is <25%. This study is summarized below:

| Summary Of In-House Repeatability | | | | | | | | | | |
|-----------------------------------|-------------|---------------|------------|------|------------|------|-------------|------|-------|-------|
| Panel<br>Member | Sample<br>N | Mean<br>AU/mL | Within-Run | | Within-Day | | Between-Run | | Total | |
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Negative 1 | 48 | 0.08 | 0.003 | 3.8% | 0.006 | 8.0% | 0.006 | 6.8% | 0.011 | 13.1% |
| Negative 2 | 48 | 0.12 | 0.005 | 4.0% | 0.003 | 6.5% | 0.009 | 7.0% | 0.013 | 10.5% |
| Negative 3 | 48 | 0.50 | 0.017 | 3.6% | 0.034 | 9.5% | 0.020 | 3.9% | 0.045 | 9.0% |
| High Negative 1 | 48 | 0.75 | 0.057 | 7.6% | 0.030 | 8.6% | 0.019 | 2.5% | 0.058 | 7.7% |
| High Negative 2 | 48 | 0.72 | 0.046 | 5.4% | 0.014 | 7.1% | 0.015 | 2.0% | 0.052 | 7.2% |

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| High Negative 3 | 48 | 0.74 | 0.015 | 2.0% | 0.018 | 4.7% | 0.011 | 1.4% | 0.034 | 4.5% |
|----------------------|----|------|-------|------|-------|-------|-------|-------|-------|-------|
| Near Cut-off 1 | 48 | 0.92 | 0.028 | 3.0% | 0.025 | 5.0% | 0.036 | 3.9% | 0.056 | 6.1% |
| Near Cut-off 2 | 48 | 1.04 | 0.022 | 2.1% | 0.014 | 3.9% | 0.033 | 3.1% | 0.045 | 4.3% |
| Near Cut-off 3 | 48 | 0.95 | 0.037 | 3.9% | 0.025 | 6.4% | 0.036 | 3.8% | 0.061 | 6.4% |
| Low Positive 1 | 48 | 1.48 | 0.029 | 2.0% | 0.029 | 3.9% | 0.014 | 0.9% | 0.058 | 3.9% |
| Low Positive 2 | 48 | 1.43 | 0.026 | 1.8% | 0.020 | 2.5% | 0.017 | 1.2% | 0.050 | 3.5% |
| Low Positive 3 | 48 | 1.65 | 0.027 | 1.6% | 0.037 | 4.2% | 0.018 | 1.1% | 0.078 | 4.7% |
| High Positive 1 | 48 | 5.43 | 0.131 | 2.4% | 0.154 | 3.8% | 0.27 | 5.0% | 0.38 | 7.0% |
| High Positive 2 | 48 | 4.85 | 0.110 | 2.3% | 0.176 | 3.6% | 0.17 | 3.6% | 0.29 | 6.0% |
| High Positive 3 | 48 | 4.74 | 0.136 | 2.8% | 0.189 | 4.9% | 0.17 | 3.5% | 4.74 | 5.2% |
| Non-Reactive Control | 48 | 0.13 | 0.004 | 3.3% | 0.008 | 6.23% | 0.007 | 5.39% | 0.010 | 8.2% |
| Reactive Control 1 | 48 | 5.63 | 0.049 | 7.5% | 5.5 | 5.15% | 0.29 | 5.11% | 0.42 | 7.51% |

#### Reproducibility

Reproducibility was evaluated internally and at two external clinical sites. The study was conducted as follows: fifteen samples were identified and/or prepared (by Zeus Scientific, Inc.) for use in the study based upon their activity on the assay. Three samples each were selected that were negative, high negative, near cut-off, low positive and high positive. To assess reproducibility, on each day of testing, each sample was diluted twice and each dilution was run in triplicate. This was repeated in a second run by a second technologist resulting in twelve results per day. This was repeated for five days at each site and the resulting data used. Reproducibility is considered acceptable for the reactive samples if the Total CV is <15% and if the Total CV for the borderline and positive samples do not congregate at the high end of acceptability but show a spread of results of at least 3%. The reproducibility for the negative sample is considered acceptable if the Total CV is <50% and shows no change in the qualitative outcome.

| Panel<br>Member | Sample<br>N | Mean<br>Index value | Within-Run | | Within-Day | | Between-Run | | Between-Site | | Total | |
|----------------------|-------------|---------------------|------------|-------|------------|-------|-------------|-------|--------------|-------|-------|-------|
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Negative 1 | 180 | 0.06 | 0.01 | 14.7% | 0.01 | 19.3% | 0.01 | 10.7% | 0.02 | 29.1% | 0.02 | 36.6% |
| Negative 2 | 180 | 0.08 | 0.01 | 9.7% | 0.01 | 12.2% | 0.01 | 8.3% | 0.01 | 14.6% | 0.01 | 15.5% |
| Negative 3 | 180 | 0.31 | 0.03 | 8.6% | 0.03 | 10.7% | 0.02 | 6.6% | 0.04 | 12.7% | 0.04 | 13.3% |
| High Negative 1 | 180 | 0.80 | 0.04 | 5.1% | 0.05 | 6.3% | 0.03 | 4.2% | 0.06 | 7.3% | 0.06 | 7.3% |
| High Negative 2 | 180 | 0.74 | 0.04 | 5.1% | 0.04 | 5.8% | 0.02 | 3.0% | 0.05 | 7.0% | 0.05 | 7.4% |
| High Negative 3 | 180 | 0.76 | 0.04 | 5.0% | 0.04 | 5.6% | 0.21 | 2.7% | 0.05 | 6.5% | 0.05 | 7.2% |
| Near Cut-off 1 | 180 | 1.05 | 0.07 | 6.3% | 0.07 | 7.3% | 0.03 | 3.6% | 0.09 | 9.0% | 0.11 | 10.3% |
| Near Cut-off 2 | 180 | 1.13 | 0.05 | 4.7% | 0.06 | 5.4% | 0.04 | 3.1% | 0.07 | 6.0% | 0.08 | 6.7% |
| Near Cut-off 3 | 180 | 0.95 | 0.05 | 5.6% | 0.07 | 6.7% | 0.04 | 4.0% | 0.08 | 8.5% | 0.09 | 9.9% |
| Low Positive 1 | 180 | 1.45 | 0.09 | 6.2% | 0.11 | 7.6% | 0.06 | 4.4% | 0.13 | 8.9% | 0.14 | 9.6% |
| Low Positive 2 | 180 | 1.77 | 0.11 | 5.9% | 0.14 | 7.8% | 0.10 | 5.8% | 0.15 | 8.3% | 0.16 | 9.2% |
| Low Positive 3 | 180 | 1.93 | 0.14 | 7.1% | 0.17 | 8.9% | 0.12 | 5.9% | 0.19 | 9.7% | 0.21 | 10.7% |
| Positive | 180 | 3.6 | 0.20 | 5.7% | 0.22 | 6.2% | 0.10 | 2.8% | 0.30 | 8.4% | 0.37 | 10.2% |
| Positive 2 | 180 | 3.1 | 0.20 | 6.2% | 0.22 | 7.3% | 0.13 | 4.4% | 0.28 | 9.0% | 0.34 | 10.9% |
| Positive 3 | 180 | 3.1 | 0.18 | 5.7% | 0.22 | 6.9% | 0.16 | 4.9% | 0.26 | 8.4% | 0.31 | 10.2% |
| Non-Reactive Control | 180 | 0.09 | 0.01 | 10.6% | 0.01 | 12.8% | 0.01 | 6.2% | 0.01 | 15.9% | 0.02 | 21.0% |
| Reactive Control | 180 | 3.9 | 0.16 | 4.1% | 0.12 | 5.1% | 0.14 | 3.5% | 0.22 | 5.7% | 0.02 | 5.9% |

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Image /page/11/Picture/1 description: The image shows the seal of the Department of Health & Human Services (HHS) of the United States. The seal features a stylized eagle with three legs, representing the department's mission to protect the health of all Americans and provide essential human services. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" is arranged in a circular pattern around the eagle.

Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993

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

FEB 10 201

Re: k102283 Trade/Device Name: Treponema pallidum IgG ELISA Test System Regulation Number: 21CFR §866.3830 Regulation Name: Treponema pallidum treponemal test reagents. Regulatory Class: Class II Product Code: LIP Dated: January 25, 2011 Received: January 26, 2011

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 the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

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

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

Page 2 - Ewa Nadolczak

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 Part 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at [phone]. 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 [phone] or [phone] or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours.

Sally A. Hojvat, M.S., Ph.D.

Sally A. Hoivat, M.Sc . Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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

## Indications for Use

510(k) Number: K102283

Device Name: ZEUS ELISA Treponema pallidum IgG Test System

Indications for Use:

The ZEUS ELISA Treponema pallidum IgG Test System is intended for the qualitative detection of specific IgG class antibodies to T. pallidum in human serum. The test may be used in conjunction with non treponemal testing and clinical findings to provide serological evidence of infection with T. pallidum. This test is for in vitro diagnostic use only.

This test is not intended for screening blood or plasma donors.

Prescription Use X AND/OR (Part 21 CFR 801 Subpart D)

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)

Leeddi m. Poole
Division Sign-Off

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Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K102283