IS H. PYLORI IGG ELISA TEST SYSTEM

K990462 · Columbia Bioscience, Inc. · LYR · Sep 3, 1999 · Microbiology

Device Facts

• Record ID: K990462
• Device Name: IS H. PYLORI IGG ELISA TEST SYSTEM
• Applicant: Columbia Bioscience, Inc.
• Product Code: LYR · Microbiology
• Decision Date: Sep 3, 1999
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3110
• Device Class: Class 1

Intended Use

For the qualitative determination of IgG antibodies to Helicobactor pylori antigen in human sera by indirect enzyme immunoassay. The H.pylori IgG assay may be used as an aid in the diagnosis of H. pylori infection in adult patients with gastrointestinal symptoms. The test can be performed either manually or in conjunction with the MAGO® PLUS Automated EIA Processor.

Device Story

The H. pylori IgG ELISA Kit is an enzyme-linked immunosorbent assay (ELISA) for detecting IgG antibodies to H. pylori antigen in human serum. Purified H. pylori antigen is bound to microwells; patient sera, calibrators, and controls are added and incubated. If present, anti-H. pylori IgG antibodies bind to the antigen. After washing, horseradish peroxidase-labeled anti-human IgG conjugate is added, followed by substrate. The resulting end product absorbance is measured spectrophotometrically at 450 nm (reference 600-630 nm). Absorbance is directly proportional to IgG concentration. The test is performed manually or using the MAGO® PLUS Automated EIA Processor in clinical laboratory settings. Results assist healthcare providers in diagnosing H. pylori infection in symptomatic adults.

Clinical Evidence

Performance evaluated using 249 retrospective patient sera characterized by biopsy (culture, stain, CLO). Sensitivity was 94.9% (112/118; 95% CI: 89.3-98.1%) and specificity was 90.4% (113/125; 95% CI: 83.8-94.9%). Overall agreement with biopsy was 92.6%. Relative agreement with another commercial ELISA was 99.6% (238/239). Precision studies (intra- and inter-assay) were conducted at three sites; manual and MAGO® PLUS automated procedures showed high correlation (r=0.993).

Technological Characteristics

Enzyme-linked immunosorbent assay (ELISA) using purified H. pylori antigen bound to microwells. Detection via horseradish peroxidase-labeled anti-human IgG conjugate and spectrophotometric measurement at 450 nm. Compatible with manual or automated (MAGO® PLUS) processing.

Indications for Use

Indicated for qualitative detection of IgG antibodies to H. pylori in human serum as an aid in diagnosing H. pylori infection in adult patients with gastrointestinal symptoms.

Regulatory Classification

Identification

Campylobacter fetus serological reagents are devices that consist of antisera conjugated with a fluorescent dye used to identify Campylobacter fetus from clinical specimens or cultured isolates derived from clinical specimens. The identification aids in the diagnosis of diseases caused by this bacterium and provides epidemiological information on these diseases. Campylobacter fetus is a frequent cause of abortion in sheep and cattle and is sometimes responsible for endocarditis (inflammation of certain membranes of the heart) and enteritis (inflammation of the intestines) in humans.

Predicate Devices

• Wampole H. pylori IgG ELISA

Submission Summary (Full Text)

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K990462

# 510k Summary of Safety and Effectiveness

# SEP , 3 1999

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is:

### Applicant Information:

| Date Prepared: | |
|----------------|------------------------------------------------------|
| Name: | Columbia Bioscience, Inc. |
| Address: | 8775 M Centre Park Drive, #559<br>Columbia, MD 21045 |

| Contact Person: | Norman Jenkins |
|-----------------|----------------|
| PhoneNumber. | 410-995-1278 |
| Fax Number. | 410-995-0508 |

#### Device Information:

| Trade Name: | Image: Logo H. pylori IgG ELISA Kit |
|----------------------|-------------------------------------|
| Common Name. | H. pylori IgG EIA Test |
| Classification Name; | H. pylori IgG Serological Reagent |

# Equivalent Device:

Wampole H. pylori IgG ELISA

Device Description: The & H. pylori IgG ELISA Kit is an enzyme-linked immunosorbent assay (ELISA) for the detection of IgG antibodies to H. pylori antigen in human serum.

Intended Use: For the qualitative determination of IgG antibodies to Helicobactor pylori antigen in human sera by indirect enzyme immunoasay. The H.pylori IgG assay may be used as an aid in the diagnosis of H. pylori infection in adult patients with gastrointestinal symptoms. The test can be performed either manually or in conjunction with the MAGO® PLUS Automated EIA Processor.

### Principle of Procedure:

Purified H. pylori antigen is bound to microwells. Diluted patient sera, Cut-Off Calibrator and controls are placed in the microwells and incubated. Anti-H. pylori IgG antibodies, if present, will bind to the antigen forming antibody complexes. Residual sample is eliminated by aspirating and washing. Conjugate (norseradish peroxidase-labeled anti-human IgG) is added and will bind to these complexes. Unbound conjugate is removed by aspiration and washing. Substrate is then added and incubated. In the presence of bound enzyme the substrate is converted to an end product. The absorbance of this end product can be read spectrophotometrically at 450 nm (reference 600-630 nm) and is directly proportional to the concentration of IgG antibodies to H. pylori present in the sample.

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# Performance Characteristics

# A. Sensitivity and Specificity Using Characterized Sera

Frozen retrospective sera from two hundred forty-nine patients were characterized using biopsy with culture, stain and CLO results for H.pylori. Based on the results of this testing, the patient sera were characterized as follows :

- * 121 sera were characterized as positive. These were positive for H. pylori by biopsy.
- * 128 sera were characterized as negative. These were negative for H. pylori by biopsy.

The sera were tested on the 1s-H, pylori IgG Test Kit at a clinical commercial laboratory. The data is summarized in Table 2.

# TABLE 2

#### Is-H. pylori IgG

| | | POSITIVE | *EQUIVOCAL | NEGATIVE | |
|-----------------------------|---------------------------------------------------|----------|-------------------------------------------------------|--------------------------------------------------|--|
| H. pylori | POSITIVE | 112 | 3 | 6 | |
| Clinical Status<br>(Biopsy) | INEGATIVE | 12 | 3 | 113 | |
| | Sensitivity<br>Specificity<br>Overall Agreement = | ll | 112/118 = 94.9%<br>113/125 = 90.4%<br>225/243 = 92.6% | 95% Cl<br>89.3-98.1%<br>83.8-94.9%<br>88.5-95.6% | |

*Equivocal results were excluded from the above calculations.

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# B. Relative Agreement Versus Another ELISA

Frozen retrospective sera from two hundred forty-nine patients (same samples from Table 2) were tested at a clinical commercial laboratory using the 1s-H, pylori 1gG Test Kit and another commercially available kit for H. pylori IgG antibodies. The data in Table 3 illustrates the relative aggrement of the 1s-H. pylori lgG Test Kit versus another commercial ELISA.

#### TABLE 3

### Is-H. pylori IgG

Another ELISA

| | POSITIVE | * EQUIVOCAL | NEGATIVE |
|------------|----------|-------------|----------|
| POSITIVE | 122 | 3 | 0 |
| *EQUIVOCAL | 1 | 1 | 3 |
| NEGATIVE | 1 | 2 | 116 |

· Of the 125 sera positive on the alternate ELISAtested, 122 were positive for Is-H. pylori IgG, none were negative, and 3 were equivocal

· Of the 119 sera negative on the alternate ELISAtested, 1 was positive for Is-H. pylori IgG, 116 were negative, and 2 were equivocal

· Of the 5 sera equivocal on the alternate ELISAtested, 1 was positive for Is-H, pylori IgG, 1 was negative, and 3 were equivocal

- · Overall Relative Agreement = 238/239 = 99.6%
* Equivocal results were excluded from calculations

NOTE : Please be advised that 'relative' relers to the comparison of the assay's results to that of a similar assay. There was not an attempt to correlate the assay's results with disease presence or absence. No judgment can be made on the comparison's accuracy to predict disease.

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# C. Precision

To determine the precision of the Is-H. pyleri IgG Test Kit, four positive sera were assayed ten times each in thee different runs at three different sites included: the manufacturer, a research & development laboratory, and a clinical commercial laboratory. The intra- and interassay precision obtained at each site is shown in Tables 4,5, and 6.

| SERUM | | INTRA-ASSAY RUN 1 | | INTRA-ASSAY RUN 2 | | INTRA-ASSAY RUN 3 | | INTERASSAY | |
|---------|---------------|-------------------|---------------|-------------------|---------------|-------------------|---------------|------------|--|
| | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | |
| A (POS) | 1.51 | 3.85 | 1.47 | 5.05 | 1.45 | 4.97 | 1.47 | 4.80 | |
| B (POS) | 2.60 | 2.16 | 2.58 | 4.20 | 2.59 | 3.69 | 2.59 | 3.35 | |
| C (POS) | 2.23 | 3.66 | 2.23 | 3.65 | 2.20 | 2.98 | 2.22 | 3.39 | |
| D (POS) | 1.73 | 3.82 | 1.72 | 2.19 | 1.70 | 3.57 | 1.71 | 3.24 | |
| E (NEG) | 0.19 | 10.81 | 0.16 | 13.54 | 0.14 | 36.64 | 0.17 | 23.60 | |
| F (NEG) | 0.36 | 37.22 | 0.32 | 16.08 | 0.34 | 11.54 | 0.34 | 24.91 | |

TABLE 4 : Site #1 - Intra-Assay and Interassay Precision

TABLE 5 : Site #2- Intra-Assay and Interassay Precision

| SERUM | INTRA-ASSAY RUN 1 | | INTRA-ASSAY RUN 2 | | INTRA-ASSAY RUN 3 | | INTERASSAY | |
|---------|-------------------|-------|-------------------|-------|-------------------|-------|---------------|-------|
| | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% |
| A (POS) | 1.34 | 4.92 | 1.32 | 5.00 | 1.38 | 4.39 | 1.35 | 4.93 |
| B (POS) | 2.25 | 5.85 | 2.27 | 5.17 | 2.45 | 6.18 | 2.33 | 6.78 |
| C (POS) | 1.97 | 3.40 | 1.96 | 4.24 | 2.11 | 4.83 | 2.01 | 5.41 |
| D (POS) | 1.46 | 6.27 | 1.45 | 4.52 | 1.62 | 4.02 | 1.51 | 6.93 |
| E (NEG) | 0.18 | 12.48 | 0.17 | 11.70 | 0.22 | 10.28 | 0.19 | 15.39 |
| F (NEG) | 0.40 | 11.01 | 0.35 | 20.01 | 0.47 | 12.50 | 0.41 | 18.56 |

TABLE 6 : Site #3 - Intra-assay and Interassay Precision

| SERUM | INTRA-ASSAY RUN 1 | | INTRA-ASSAY RUN 2 | | INTRA-ASSAY RUN 3 | | INTERASSAY | |
|---------|-------------------|-------|-------------------|-------|-------------------|-------|---------------|-------|
| | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% |
| A (POS) | 1.41 | 7.59 | 1.30 | 5.70 | 1.27 | 4.60 | 1.33 | 7.64 |
| B (POS) | 2.54 | 4.41 | 2.30 | 3.74 | 2.19 | 2.52 | 2.34 | 7.31 |
| C (POS) | 2.13 | 5.91 | 1.96 | 4.89 | 1.90 | 3.43 | 2.00 | 6.94 |
| D (POS) | 1.56 | 6.16 | 1.50 | 4.59 | 1.45 | 2.08 | 1.50 | 5.42 |
| E (NEG) | 0.16 | 19.99 | 0.18 | 6.38 | 0.16 | 6.25 | 0.17 | 13.77 |
| F (NEG) | 0.34 | 10.92 | 0.34 | 13.65 | 0.27 | 15.74 | 0.32 | 16.60 |

### D. Correlation of Manual and MAGO Plus Results

The Is-H. pylori IgG Test Kit has been developed for automated as . To demonstrate the equivalence of the manual and MAGO Plus procedures, the results of 100 serum samples tested by both methods were plotted. A scattergram and regression line of the results obtained with 95% confidence intervals is shown in Figure 3. The data indicate good correlation with a Pearson Correlation Coefficient of 0.993.

Image /page/3/Figure/10 description: The image is a scatter plot that shows the correlation between MANUAL Index Values and MAGO PLUS Index Values. The x-axis represents the MANUAL Index Values, ranging from 0.0 to 4.0, while the y-axis represents the MAGO PLUS Index Values, ranging from -0.5 to 5.0. The scatter plot shows a strong positive correlation between the two variables, with a correlation coefficient of r = 0.993. A regression line is plotted through the data points.

FIGURE 3 : Manual and MAGO Plus Result Correlation

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# E. MAGO Plus Precision

The precision of the assay when performed on the MAGO Plus Automated EIA Processor was determined by assaying six sera ten times each in three different runs. Table 7 shows the intra-and interassay precision obtained using the MAGO Plus.

| SERUM | INTRA-ASSAY RUN 1 | | INTRA-ASSAY RUN 2 | | INTRA-ASSAY RUN 3 | | INTERASSAY | |
|---------|-------------------|-------|-------------------|-------|-------------------|-------|---------------|-------|
| | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% |
| A (POS) | 1.19 | 8.36 | 1.02 | 15.19 | 1.25 | 5.66 | 1.15 | 12.83 |
| B (POS) | 2.12 | 4.87 | 2.20 | 9.34 | 2.27 | 2.97 | 2.20 | 6.71 |
| C (POS) | 1.97 | 4.18 | 1.89 | 5.82 | 1.90 | 4.96 | 1.92 | 5.19 |
| D (POS) | 1.37 | 6.01 | 1.45 | 4.88 | 1.54 | 7.62 | 1.45 | 7.82 |
| E (NEG) | 0.13 | 37.16 | 0.12 | 35.14 | 0.20 | 62.36 | 0.15 | 57.40 |
| F (NEG) | 0.32 | 24.65 | 0.34 | 24.80 | 0.34 | 15.19 | 0.33 | 21.33 |

TABLE 7 : Site #2- Intra-Assay and Interassay Precision - MAGO Plus

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Image /page/5/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular arrangement of text surrounding a stylized symbol. The text reads "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" in a circular fashion. The symbol in the center appears to be an abstract representation of a bird or a stylized human figure.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

SEP 3 1999

Mr. William Boteler Immunoprobe, Inc. 1306F Bailes Lane Frederick, Maryland 21701

Re: K990462 Trade Name: Helicobactor pylori IgG ELISA Regulatory Class: I Product Code: LYR Dated: June 15, 1999 Received: June 16, 1999

#### Dear Mr. Boteler:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we 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). 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 either class II (Special Controls) or class III (Premarket Approval), 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 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic OS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

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Page 2

Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at [phone].

This letter will allow you to begin marketing your device as described in your 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 additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at [phone]. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll free number [phone] or at [phone] or at its internet address "http://www.fda.gov/cdrh/dsmamain.html"

Sincerely yours,

Steven Butman

Steven I. Gutman, M.D., M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure

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510(k) Number:

Device Name: 20 H. pylori IgG ELISA

Indications For Use: For the qualitative determination of IgG antibodies to Helicobactor pylori antigen in human sera by indirect enzyme immunoassay. The H.pylori IgG assay may be used as an aid in the diagnosis of H. pylori infection in adult patients with gastrointestinal symptoms. The test can be performed either manually or in conjunction with the MAGO® PLUS Automated EIA Processor.

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Prescription Use (Per 21 CFR 801.109) OR

w===========================================================================================================================================================================

Over-The Counter Use (Optional Format 1-2-96)

Woody Dubaes

(Division Sign Off) Division of Clinical Laboratory Devices 510(k) Number_19