WIELISA ANTI-GBM, ANCA SCREENING KIT TEST SYSTEM

K981750 · Wieslab AB · MOB · Jul 22, 1998 · Immunology

Device Facts

• Record ID: K981750
• Device Name: WIELISA ANTI-GBM, ANCA SCREENING KIT TEST SYSTEM
• Applicant: Wieslab AB
• Product Code: MOB · Immunology
• Decision Date: Jul 22, 1998
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.5660
• Device Class: Class 2

Intended Use

The Wielisa Anti-GBM, ANCA Screening Test Kit is an enzymelinked immunosorbent assay (ELISA) for the qualitative detection of antibodies to glomerular basement membrane (GBM), Proteinase-3 (PR-3) and Myeloperoxidase (MPO) in human sera. The assay is used to detect antibodies in a single serum specimen. The results of the assay are to be used as an aid to the diagnosis of reno-pulmonary syndromes and rapidly progressive glomerulonephritis, especially Goodpasture syndrome (GP), Wegener's granulomatosis (WG) and microcopic polyangiitis (MP). The assay is intended for use in patients with signs and symptoms consistent with GP, WG, and MP. It is not intended for screening a healthy population. A positive result should always be confirmed by a semi-quantitative assay.

Device Story

Wielisa Anti-GBM, ANCA Screening Test Kit is an ELISA for qualitative detection of antibodies to GBM, PR-3, and MPO in human serum. Microtiter wells are coated with purified antigens; patient serum is added; specific antibodies bind to antigens. Unbound components are washed away; alkaline phosphatase-labeled anti-human IgG conjugate is added. After further washing, substrate is added; color intensity (optical density) correlates to bound antibody concentration. Results are calculated as a ratio to negative control. Used in clinical laboratories by trained personnel to aid diagnosis of reno-pulmonary syndromes and glomerulonephritis. Output is a qualitative result (negative, equivocal, positive) used by physicians to guide clinical decision-making; positive results require semi-quantitative confirmation. Benefits include rapid screening for specific autoimmune conditions.

Clinical Evidence

Clinical evidence based on 326 frozen retrospective sera. Sensitivity: PR-3 (WG 92.9%, MP 51.2%), MPO (WG 9.8%, MP 46.5%), anti-GBM (GP 100%). Specificity: PR-3 (100% across all groups), MPO (SLE 82.8%, RA 100%, NS 100%), anti-GBM (100% across all groups). Relative sensitivity/specificity compared to semi-quantitative ELISA also reported (e.g., PR-3 relative sensitivity 98.3%).

Technological Characteristics

ELISA-based immunoassay. Microtiter strips coated with purified human neutrophil-derived PR-3, MPO, and bovine-derived GBM antigens. Detection via alkaline phosphatase-labeled goat anti-human IgG conjugate and substrate solution. Measurement via optical density (OD) ratio. Standalone manual or automated plate reader format.

Indications for Use

Indicated for qualitative detection of anti-GBM, PR-3, and MPO antibodies in human sera to aid diagnosis of reno-pulmonary syndromes and rapidly progressive glomerulonephritis (Goodpasture syndrome, Wegener's granulomatosis, microscopic polyangiitis) in symptomatic patients. Not for healthy population screening. Positive results require semi-quantitative confirmation.

Regulatory Classification

Identification

A multiple autoantibodies immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoantibodies (antibodies produced against the body's own tissues) in serum and other body fluids. Measurement of multiple autoantibodies aids in the diagnosis of autoimmune disorders (disease produced when the body's own tissues are injured by autoantibodies).

Predicate Devices

• Wielisa PR-3 ANCA ELISA Kit
• Wielisa MPO ANCA ELISA Kit
• Wielisa anti-GBM ELISA Kit

Submission Summary (Full Text)

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L

K981750

Summary of Safety and Effectiveness Information GBM, ANCA Screening ELISA Test Kit

Wieslab AB Ideon Research Park S-223 70 Lund Sweden Contact person: Dr. Jorgen Wieslander Telephone: [phone] Date of preparation: May 1, 1998

Description of Device: The Wielisa Anti-GBM, ANCA Screening Test Kit is an II. enzyme-linked immunosorbent assay (ELISA) for the qualitative detection of antibodies to glomerular basement membrane (GBM), Proteinase-3 (PR-3) and Myeloperoxidase (MPO) in human sera. The assay is used to detect antibodies in a single serum specimen. The results of the assay are to be used as an aid to the diagnosis of reno-pulmonary syndromes and rapidly progressive glomerulonephritis, especially Goodpasture syndrome (GP). Wegener's granulomatosis (WG) and microcopic polyangiitis (MP). The assay is intended for use in patients with signs and symptoms consistent with GP, WG, and MP. It is not intended for screening a healthy population. A positive result should always be confirmed by a semi-quantitative assay.

The wells of the microtiter strips are coated with purified proteinase 3 (Human Neutrophil source). MPO (Human Neutrophil source) and GBM (Bovine source) antigen. During the first incubation, specific antibodies in diluted serum, will bind to the antigen coating.

The wells are then washed to remove unbound antibodies and other components. A coniugate of alkaline phosphatase-labeled (Goat) antibodies to human IgG binds to the antibodies in the wells in this second incubation.

After a further washing step, detection of specific antibodies is obtained by incubation with substrate solution. The amount of bound antibodies correlates to the color intensity and is measured in terms of absorbance (optical density (OD)). The absorbance is then calculated and the results are given as a ratio to the negative control.

#### III. Predicate Device

The GBM, ANCA Screening test is substantially equivalent to the Wielisa PR-3 ANCA ELISA Kit , the Wielisa MPO ANCA ELISA Kit and the Wielisa anti-GBM ELISA Kit. Equivalence is demonstrated by the following comparative results:

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| Control and<br>Disease groups | Total | Negative <3 | | | Equivocal 3-4 | | | Positive >4 | | |
|-------------------------------|-------|-------------|-----|-----|---------------|-----|-----|-------------|-----|-----|
| | | GBM | PR3 | MPO | GBM | PR3 | MPO | GBM | PR3 | MPO |
| Blood donors: (NS) | 131 | 128 | 131 | 127 | 3 | 0 | 4 | 0 | 0 | 0 |
| WG: | 42 | - | 3 | 37 | - | 0 | 1 | - | 39 | 4 |
| MP: | 43 | - | 20 | 23 | - | 2 | 0 | - | 21 | 20 |
| SLE: | 31 | 31 | 31 | 24 | 0 | 0 | 2 | 0 | 0 | 5* |
| RA: | 41 | 41 | 41 | 40 | 0 | 0 | 1 | 0 | 0 | 0 |
| GP: | 38 | 0 | - | - | 0 | - | - | 38 | - | - |

Table 1. Clinical sensitivity and specificity. A total of 326 frozen retrospective sera with clinical characterisation were assayed. The following table summarises the results

MP = microscopic polyangiitis RA = rheumatoid arthritis WG = Wegener's granulomatosis,

SLE = systemic lupus erythematosus GP = Goodpasture syndrome

* All samples were positive in semi-quantitative MPO-ELISA.

#### Clinical sensitivity (Equivocal samples are not included in the calculation)

| PR3-ANCA: | WG = 39/42 = 92.9 % | 95% CI = 84.9 – 100% |
|-----------|---------------------|----------------------|
| | MP = 21/41 = 51.2 % | 95% CI = 35.6-66.8% |
| MPO-ANCA: | WG = 4/41 = 9.8 % | 95% CI = 4.9 – 19.0% |
| | MP = 20/43 = 46.5 % | 95% CI = 31.3-61.7% |
| Anti-GBM: | GP = 38/38 = 100 % | 95% CI = 92.2 – 100% |

#### Clinical specificity (Equivocal samples are not included in the calculation)

| PR3-ANCA: | SLE = 31/31 = 100 %<br>RA = 41/41 = 100 %<br>NS = 131/131 = 100 % | 95% CI = 90.4 – 100%<br>95% CI = 92.7 - 100%<br>95% CI = 97.7 - 100% |
|-----------|--------------------------------------------------------------------|-----------------------------------------------------------------------|
| MPO-ANCA: | SLE = 24/29 = 82.8 %<br>RA = 40/40 = 100 %<br>NS = 127/127 = 100 % | 95% CI = 68.7 - 96.8%<br>95% CI = 92.6 - 100%<br>95% CI = 97.6 - 100% |
| Anti-GBM: | SLE = 31/31 = 100 %<br>RA = 41/41 = 100 %<br>NS = 128/128 = 100 % | 95% CI = 90.4 -100%<br>95% CI = 92.7 - 100%<br>95% CI = 97.6 – 100% |

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Table 2. Relative sensitivity and specificity of the Wielisa anti-GBM, ANCA screen kit compared to an alternative semi-quantitative ELISA. A total of 216 frozen retrospective sera were assayed on the Wielisa anti-GBM, ANCA screen kit and a semi-quantitative ELISA for PR-3 and MPO. Also, a total of 169 frozen retrospective sera were assayed on the Wielisa anti-GBM, ANCA screen kit and an semiquantitative ELISA for anti-GBM. The following table summarises the results.

| Semi-quantitative<br>ELISA | Negative <3 | | | Equivocal | | | Positive >4 | | | |
|----------------------------|-------------|-----|-----|-----------|-----|-----|-------------|-----|-----|----|
| | GBM | PR3 | MPO | GBM | PR3 | MPO | GBM | PR3 | MPO | |
| MPO-ANCA | Positive | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 23 |
| PR3-ANCA | Positive | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 59 | 0 |
| Anti-GBM | Positive | 0 | - | - | 0 | - | - | 37 | - | - |
| | Negative | 128 | 152 | 182 | 3 | 2 | 4 | 0 | 0 | 0 |
| | Equivocal | 0 | 1 | 4 | 0 | 0 | 1 | 1 | 1 | 1 |
| | Total | 128 | 154 | 187 | 3 | 2 | 5 | 38 | 60 | 24 |

Relative sensitivity (Equivocal samples are not included in the calculation)

Relative sensitivity PR3-ANCA = 59/60 = 98.3% 95% CI = 95.0 - 100% Relative sensitivity MPO-ANCA = 23/24 = 95.8% 95% CI = 87.7 - 100% Relative sensitivity anti-GBM = 37/37 = 100 % 95% CI = 92.0 - 100%

#### Relative specificity (Equivocal samples are not included in the calculation)

Relative specificity PR3-ANCA = 152/152 = 100 % 95% CI = 98.0 - 100% Relative specificity MPO-ANCA= 182/182 = 100 % 95% CI = 98.4 - 100% Relative specificity anti-GBM = 128/128 = 100 % 95% CI = 97,7 - 100%

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Table 3. Batch to batch variation was determined by testing three different samples. Results were obtained for 4 different batches.

| PR3 | OD ratio | | Sample Mean SD CV% Sample Mean SD CV% Sample Mean SD CV%<br>MPO | OD ratio | | GBM | OD ratio | all and the controlled on the consideration of the comments of the comments of the comments of the comments of the many of the many of the many of the many of the many of the |
|-----|----------|--|--------------------------------------------------------------------------|----------|--|--------------------------------------------------|----------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| 2 | | | 37.5 × 3.8 × 10 × 10 × 3 × 3 × 16.8 × × 2.2 × 13 × 13 × × 1 × 1 × × | | | | 21.8 | |
| | | | 24.3 23.6 3.6 15 15 15 6 1 | | | 31.5 1.5 1.9 1.9 6 6 2 4 4 29.0 29.0 2.2 2.2 4 4 | | |
| 8 | | | 35.5 % 2.4 % 777 % % 9 % 28.3 % % 1.5 % % 5 % % 7 % % 33.8 % % 2.4 % 7 % | | | | | |

Table 4. Inter-assay precision was determined by testing one sample. Results were obtained for six different runs.

| Sample | Mean<br>OD ratio | SD | CV % | Sample<br>MPO | Mean<br>OD ratio | SD | CV % | Sample<br>GBM | Mean<br>OD ratio | SD | CV % |
|--------|------------------|-----|------|---------------|------------------|------|------|---------------|------------------|------|------|
| PK | 27.3 | 2.9 | 11 | PK | 17.3 | 3.8 | 21 | PK | 16.5 | 1.1 | 6 |
| K5 | 12.1 | 1.2 | 10 | K6 | 16.5 | 0.84 | 5 | K4 | 3.6 | 0.25 | 7 |

Table 5. Intra-assay precision was determined by testing one sample in 22 wells.

| Sample<br>PR3 | Mean<br>OD | SD | CV % | Sample<br>MPO | Mean<br>OD | SD | CV % | Sample<br>GBM | Mean<br>OD | SD | CV% |
|---------------|------------|------|------|---------------|------------|------|------|---------------|------------|------|-----|
| PK | 1.3 | 0.07 | 6 | PK | 1.8 | 0.06 | 3 | PK | 1.2 | 0.19 | 16 |
| K5 | 1.3 | 0.06 | 5 | K6 | 1.46 | 0.07 | 5 | K4 | 0.6 | 0.03 | 6 |

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Image /page/4/Picture/2 description: The image is a black and white logo for the U.S. Department of Health & Human Services. The logo features a stylized abstract symbol resembling an eagle or bird in flight. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the symbol. The text is in all capital letters and is evenly spaced around the circle.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

## JUL 22 1998

Weislab AB c/o William L. Boteler, Jr. IMMUNO PROBE, INC. 1306 Bailes Lane, Suite F Frederick, MD 21701

Re: K981750 Trade Name: Wielisa Anti-GBM, ANCA Screening Test Kit Regulatory Class: II Product Code: DBL, MOB Dated: May 3, 1998 Received: May 18, 1998

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 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 requirement, as set forth in the Quality System Regulation (QS) for Medical Devices: General requlation (21 CFR Part 820) and that, through periodic (QS) 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 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal Laws or Requlations.

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Page 2

Under the Clinical Laboratory Improvement Amendments of 1988 (CDIA-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 Sutman

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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Page 1 of 1

# 510(k) Number: Not known K98) 750

Device Name: Wielisa Anti-GBM, ANCA Screening Test Kit

Indications For Use: The Wielisa Anti-GBM, ANCA Screening Test Kit is an enzymelinked immunosorbent assay (ELISA) for the qualitative detection of antibodies to glomerular basement membrane (GBM), Proteinase-3 (PR-3) and Myeloperoxidase (MPO) in human sera. The assay is used to detect antibodies in a single serum specimen. The results of the assay are to be used as an aid to the diagnosis of reno-pulmonary syndromes and rapidly progressive glomerulonephritis, especially Goodpasture syndrome (GP), Wegener's granulomatosis (WG) and microcopic polyangiitis (MP). The assay is intended for use in patients with signs and symptoms consistent with GP, WG, and MP. It is not intended for screening a healthy population. A positive result should always be confirmed by a semi-quantitative assay.

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Prescription Use V (Per 21 CFR 801.109)

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OR

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

(Division Sign-Off)
Division of Clinical Laboratory Devices K981,750
510(k) Number