WIESLAB CAP PR-3 ANCA

K051458 · Eurodiagnostica · MOB · Nov 22, 2005 · Immunology

Device Facts

• Record ID: K051458
• Device Name: WIESLAB CAP PR-3 ANCA
• Applicant: Eurodiagnostica
• Product Code: MOB · Immunology
• Decision Date: Nov 22, 2005
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.5660
• Device Class: Class 2

Indications for Use

The Wieslab™ Cap PR3-ANCA is an enzyme-linked immunosorbent assay (ELISA) for detection and semi-quantitation of IgG antibodies to proteinase 3 (PR3) 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 Wegener's granulomatosis. The assay is intended for use in patients with signs and symptoms consistent with WG. It is not intended for screening a healthy population. The analysis should be performed by trained laboratory professionals.

Device Story

Wieslab™ Cap PR3-ANCA is an ELISA test kit for detecting IgG antibodies to proteinase 3 (PR3) in human serum. Microtiter wells are coated with monoclonal antibody to PR3 and PR3 antigen. Patient serum is incubated in wells; specific antibodies bind to antigen. After washing, alkaline phosphatase-labeled anti-human IgG conjugate is added. Substrate solution is added; color intensity (optical density) correlates to bound antibody concentration. Performed by trained laboratory professionals in clinical settings. Results are calculated against a calibrator curve in arbitrary units. Output aids physicians in diagnosing Wegener's granulomatosis in symptomatic patients.

Clinical Evidence

Clinical performance evaluated on 363 characterized sera. Sensitivity for Wegener’s granulomatosis (WG) was 96.0% (95% CI: 83.3-99.5%). Specificity in healthy controls was 100% (95% CI: 97.0-100%), SLE 100% (95% CI: 92.0-100%), and RA 100% (95% CI: 80.5-100%). Method comparison with predicate (n=180) showed 98.3% overall agreement. Analytical precision (intra-assay CV 3-13%, inter-assay CV 4-14%) and linearity (10-320 U/mL) were established.

Technological Characteristics

ELISA-based immunological test system. Components: microtiter strips coated with monoclonal antibody to PR3 and PR3 antigen. Detection: alkaline phosphatase-labeled anti-human IgG conjugate and substrate solution. Measurement: optical density (OD) via spectrophotometry. Quantitative output in arbitrary units.

Indications for Use

Indicated for patients with signs and symptoms consistent with Wegener’s granulomatosis (WG) to aid in diagnosis. Not intended for screening healthy populations.

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

• Wieslisa® PR-3 ANCA Kit (k974167)

Submission Summary (Full Text)

{0}

1

# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number:
k051458

B. Purpose for Submission:
New Device

C. Measurand:
Anti- PR3 antibodies

D. Type of Test:
Qualitative and Semi-quantitative ELISA

E. Applicant:
Eurodiagnostica

F. Proprietary and Established Names:
Wieslab™ Cap PR-3 ANCA

G. Regulatory Information:
1. Regulation section:
21 CFR 866.5660, Multiple autoantibodies immunological test system
2. Classification:
II
3. Product codes:
MOB, Test system, antineutrophil cytoplasmic antibodies (ANCA)
4. Panel:
Immunology 82

H. Intended Use:
1. Intended use(s):
The Wieslab™ Cap PR-3 ANCA test kit is an enzyme-linked immunosorbent assay (ELISA) for detection and semi-quantitation of IgG antibodies to proteinase 3 (PR3) in human serum. 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 Wegener’s granulomatosis. The analysis should be performed by trained laboratory professionals. For In Vitro Diagnostic Use.
2. Indication(s) for use:
The Wieslab™ Cap PR-3 ANCA test kit is an enzyme-linked immunosorbent assay (ELISA) for detection and semi-quantitation of IgG antibodies to proteinase 3 (PR3) in human serum. 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 Wegener’s granulomatosis. The assay is intended for use in patients with signs and symptoms consistent with WG. It is not intended for screening a healthy population. The analysis should be performed by trained laboratory professionals.
3. Special conditions for use statement(s):
The device is for prescription use only.
4. Special instrument requirements:
Microplate reader capable of measuring OD at 405 nm.
Microplate washer (300μL volume).

{1}

2

I. Device Description:

Each device contains the following: microplate strips (red colored) coated with monoclonal anti-proteinase 3/proteinase 3 antigen; five levels calibrators (10, 40, 80, 160, 320 U/mL); positive and negative controls (human serum in diluent); wash solution concentrate; sample diluent; goat anti-human IgG alkaline phosphatase conjugate; $p$-Nitro phenyl Phosphate ($p$NPP) substrate.

J. Substantial Equivalence Information:

1. Predicate device name(s):
Wieslisa® PR-3 ANCA Kit

2. Predicate 510(k) number(s):
k974167

3. Comparison with predicate:

| Similarities | | |
| --- | --- | --- |
| Item | New Device | Predicate Device |
| Intended use | To aid in the diagnosis of Wegener’s granulomatosis (WG) | Same |
| Assay Format | Qualitative and semi-quantitative | Same |
| Enzyme-Conjugate | Alkaline Phosphatase | Same |
| Positive and Negative controls | Ready to use | Same |
| Calibrators: Five Levels | Ready to use | Same |
| Sample volume required | 5 μL | Same |
| Sample type and dilution | Serum at 1:80 | Same |
| Sample diluent | PBS | Same |
| Wash solution | 30x Concentrate | Same |
| Substrate | pNPP | Same |
| Incubation times | 30-30-60 minutes | Same |
| OD reading | 405 nm | Same |
| Platform | 96 well microtiter plates | Same |
| Differences | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Technology | Capture ELISA | Direct ELISA |
| Antigen | Purified anti-PR3 monoclonal antibody and PR3 capture complex | Purified PR3 |
| OD measurement | Within ± 5 minutes | None specified |
| Anti-PR-3 Antibody Results Interpretation | Negative: < 21 U/mL
Equivocal: 21-30 U/mL
Positive: > 30 U/mL | Negative: < 10 Units
Equivocal: 10-20 Units
Positive: > 20 Units |

K. Standard/Guidance Document Referenced (if applicable):

None provided.

L. Test Principle:

The Wieslab™ Cap PR-3 ANCA test kit is an enzyme-linked immunosorbent assay (ELISA) for detection and semi-quantitation of IgG antibodies to proteinase 3 (PR3) in human serum. The wells

{2}

of the microtiter plate strips are coated with a capture complex of monoclonal antibody to proteinase 3 and purified PR3 antigen. Antibodies specific to PR3 in diluted serum bind to the capture antigen during the first incubation. The wells are then washed to remove unbound antibodies. An enzyme labeled anti-human IgG conjugate is added to bind the capture antigen-antibody complex in the well during the second incubation step. After a further washing step, detection of the specific antibodies is obtained by incubation with substrate solution. The amount of bound antibodies correlated to the color intensity and is measured in terms of absorbance (optical density, OD). The absorbance is then calculated against the calibrator curve and the results are given in arbitrary Units/mL.

M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

The intra-assay reproducibility was determined by testing eight samples 24 times. Five samples with high anti-PR3 concentrations (102-141 U/mL) had a CV of 6-9% and three samples close to the assay cut-off (24-42 U/mL) had a CV of 3-13%. The inter-assay reproducibility was determined by testing seven samples in duplicate for four times. Three samples with high anti-PR3 concentrations (77-138 U/mL) had a CV of 6-7%. Four samples close to the assay cut-off (23-49 U/mL) had a CV of 4-14%.

b. Linearity/assay reportable range:

Three positive sera were diluted serially from neat, 1:2, 1:4, 1:8, 1:16 and 1:32 dilutions. The values were compared to log 2 of dilution by standard regression. The values indicate that the assay has a linear relationship with serum dilutions. The assay reportable range is from 10-320 U/mL.

c. Traceability, Stability, Expected values (controls, calibrators, or methods):

There is no reference standard for anti-PR3. The calibrators and controls (positive and negative) are prepared in-house and arbitrary units are assigned during the development process.

d. Detection limit:

Not applicable.

e. Analytical specificity:

Interference by endogenous substances: No data provided. The package insert states to avoid sera which are icteric, lipemic, and hemolyzed; and heat-inactivated sera should not be used.

Crossreactivity with heterophile antibodies: Since one of the capture antigen component is a monoclonal antibody, some in-house interference studies (~12000 samples) were performed. The effect was determined to be very low (0.025%). The package insert states that 'individuals receiving mouse anti-human antibodies for treatment or diagnosis, or those patients who have otherwise exposed to mouse immunoglobulin, may produce Human Anti-Mouse Antibodies (HAMA). These antibodies can interfere with assays using mouse monoclonal antibodies and may cause falsely elevated levels'.

f. Assay cut-off:

The cut-off value of &gt;30 U/mL was based on testing 120 normal blood donor sera, 17 rheumatoid arthritis (RA) samples, 46 systemic lupus erythematosus (SLE). All of the

3

{3}

normal donors were negative. One of the RA and one of the SLE samples were equivocal (23 and 27 U/mL respectively).

2. Comparison studies:

a. Method comparison with predicate device:

Testing was performed on 180 samples. The positive percent agreement was 97.6% (40/41); the negative percent agreement was 98.5% (133/135) and the Overall Agreement was 98.3% (173/176).

| | Wieslisa PR3-ANCA ELISA Kit | | | | |
| --- | --- | --- | --- | --- | --- |
| | | Positive | Equivocal* | Negative | Total |
| WiesLab™
Cap PR3-ANCA | Positive | 40 | (2)* | 2 | 42 |
| | Equivocal | 0 | 0 | (2)* | (2)* |
| | Negative | 1 | 0 | 133 | 134 |
| | Total | 41 | (2)* | 135 | 176 |

*Equivocal results were excluded in the calculation.

b. Matrix comparison:

Not applicable.

3. Clinical studies:

a. Clinical sensitivity and specificity:

The clinical sensitivity and specificity study were evaluated on 363 clinically characterized sera from patients with the following diagnosis: 50 WG; 50 microscopic polyangiitis (MP); 45 SLE; 18 RA; 80 glomerular basement membrane antibody (GBM); and 120 healthy blood donors. Sensitivity for WG was 97%. Sensitivity for MP patients was 31.2% because PR3-ANCA antibodies are only found in approximately one third of the MP patients. The overall specificity of the new device (healthy and disease controls) was 97.3%.

| N= 363 | | ELISA | | |
| --- | --- | --- | --- | --- |
| Patient Group | n= | positive | Equivocal* | negative |
| WG | 50 | 48 | 0 | 2 |
| MP | 50 | 15 | (2)* | 33 |
| Healthy controls | 120 | 0 | 0 | 120 |
| Disease controls | | | | |
| SLE | 45 | 0 | 1 | 44 |
| RA | 18 | 0 | 1 | 17 |
| GBM | 80 | 4 | 0 | 76 |

*Equivocal results were excluded from the calculation.

Sensitivity:

WG: 96.0% (48/50) 95% CI: 83.3% to 99.5%

MP: 31.2% (15/48) 95% CI: 18.7% to 46.2%

Specificity:

Healthy controls: 100% (120/120) 95% CI: 97.0% to 100%

SLE: 100% (44/44) 95% CI: 92.0% to 100%

{4}

RA: 100% (17/17) 95% CI: 80.5% to 100%
GBM: 95% (76/80) 95% CI: 97.0% to 100%)

b. Other clinical supportive data (when a. is not applicable):
Not Applicable.

4. Clinical cut-off:
Same as assay cut-off.

5. Expected values/Reference range:
Expected values in the normal population should be negative.

N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

5