IMMUGLO ANTI-ENDOMYSIAL ANTIBODY IFA
Device Facts
| Record ID | K060157 |
|---|---|
| Device Name | IMMUGLO ANTI-ENDOMYSIAL ANTIBODY IFA |
| Applicant | Immco Diagnostics, Inc. |
| Product Code | MVM · Immunology |
| Decision Date | May 25, 2006 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 866.5660 |
| Device Class | Class 2 |
Indications for Use
An indirect immunofluorescence assay (IFA) for the detection of and semi-quantitation of endomysial antibodies (EMA) in human serum to aid in the diagnosis of Celiac Disease (CD) and Dermatitis Herpetiformis (DH) in combination with other clinical and other laboratory findings.
Device Story
Indirect immunofluorescence assay (IFA) for detection/semi-quantitation of endomysial antibodies (EMA) in human serum; aids diagnosis of Celiac Disease and Dermatitis Herpetiformis. Performed in clinical laboratory settings by trained personnel. Input: human serum sample; substrate (monkey esophagus tissue). Process: serum incubated on substrate; EMA binds to endomysium; fluorescein-labeled anti-human IgG conjugate added; visualization via fluorescence microscopy. Output: qualitative/semi-quantitative presence of antibodies. Clinical utility: supports diagnosis when combined with other clinical/laboratory findings.
Clinical Evidence
No clinical data provided in the document; substantial equivalence established via bench testing and comparison to predicate devices.
Technological Characteristics
Indirect immunofluorescence assay (IFA). Utilizes monkey esophagus tissue as substrate. Detection via fluorescein-labeled anti-human IgG conjugate. Manual visualization via fluorescence microscopy. In vitro diagnostic test system.
Indications for Use
Indicated for patients suspected of having celiac disease or dermatitis herpetiformis to aid in diagnosis via detection of IgA or IgG endomysial antibodies in human serum.
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).
Related Devices
- K061408 — EUROIMMUN ENDOMYSIUM IFA: ESOPHAGUS (MONKEY) KIT · Euroimmun Us, LLC · Oct 5, 2006
- K980312 — NOVA LITE ENDOMYSIAL · Inova Diagnostics, Inc. · Apr 2, 1998
- K993034 — MEDICA INDIRECT IMMUNOFLUORESCENCE (IIF) ANTI- ENDOMYSIAL ANTIBODY (AEMA) TEST KIT, MODEL 6001-ESD · Medica, Inc. · Nov 1, 1999
Submission Summary (Full Text)
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
A. 510(k) Number:
k060157
B. Purpose for Submission:
New Devices
C. Measurand:
Anti-Endomysial Antibody
D. Type of Test:
Qualitative and semi-quantitative, indirect immunofluorescence (IFA)
E. Applicant:
IMMCO Diagnostics, Inc.
F. Proprietary and Established Names:
ImmuGlo™ Anti-Endomysial IFA
G. Regulatory Information:
1. Regulation section:
21 CFR 866.5660, Multiple Autoantibodies Immunological test system
2. Classification:
II
3. Product code:
MVM, Autoantibodies, Endomysial (Tissue Transglutaminase)
4. Panel:
Immunology 82
H. Intended Use:
1. Intended use(s):
An indirect immunofluorescence antibody test for the qualitative and semi-quantitative detection of endomysial antibodies (EMA of IgA or IgG) in human serum as an aid in the diagnosis of celiac disease and dermatitis herpetiformis in combination with other clinical and other laboratory findings.
2. Indication(s) for use:
Same as intended use.
3. Special conditions for use statement(s):
For prescription use only.
4. Special instrument requirements:
Fluorescence microscope
I. Device Description:
Each device contains the following: 8x6-well-Primate Distal Esophagus Substrate slides, EMA positive and negative controls, goat anti-human FITC IgA and IgG conjugates with Evan’s Blue Counter stain, buffered diluent, phosphate buffered saline, mounting medium, cover slips.
J. Substantial Equivalence Information:
1. Predicate device name(s):
ImmuGlo™ EMA IFA System with Primate Smooth Muscle
ImmuLisa™ tTgG IgG ELISA
2. Predicate 510(k) number:
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k912551 ImmuGlo™ EMA IFA
k040095 ImmuLisa™ tTG IgG ELISA
3. Comparison with predicate:
| Similarities | | |
| --- | --- | --- |
| Item | New Device | Predicate Device |
| Intended use | To aid in the diagnosis of Celiac Disease (CD) and dermatitis herpetiformis | Same |
| Anti-EMA IgA and IgG assay format | Qualitative and semi-quantitative | Same |
| Anti-EMA IgA and IgG Controls | Positive and Negative Controls | Same |
| Anti-EMA IgA technology | IFA | Same |
| Anti-EMA IgA (IFA) platform | 8x6 well slides | Same |
| Anti-EMA IgA (IFA) sample dilution | 1:2.5 | Same |
| Anti-EMA IgA (IFA): Incubation time | 30-30 minute | Same |
| Anti-EMA IgA (IFA): Wash Step | Two 10 minute wash | Same |
| Anti-EMA IgA (IFA) Cut-off | 2.5 Titer | Same |
| Anti-EMA IgA semi-quantitative result | End point Titer | Same |
| Differences | | |
| --- | --- | --- |
| Item | New Device | Predicate |
| Anti-EMA antigen | IgA and IgG (IFA): Primate smooth muscle from distal esophagus | IgA (IFA): Primate smooth muscle from the bladder IgG (ELISA): Recombinant human tTG |
| Anti-EMA Conjugate | FITC IgA and FITC IgG | IgA (IFA): FITC polyvalent IgG (ELISA): Alkaline phosphatase |
| Anti-EMA IgG technology | IFA | ELISA |
| Anti-EMA IgG platform | 8x6 well slides | ELISA: 12x8 Microwells |
| Anti-EMA IgG sample dilution | 1:2.5 | ELISA: 1:51 |
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| Differences | | |
| --- | --- | --- |
| Item | New Device | Predicate |
| Anti-EMA IgG: calibrators | Not Applicable | ELISA: 4 level calibrators |
| Anti-EMA IgG: substrate | Not Applicable | ELISA: pNPP |
| Anti-EMA IgG incubation times (min) | 30-30 | 30-30-30 |
| Anti-EMA IgG: Wash step | Two 10 minute wash | Two 4X wash |
| Anti-EMA IgG: reading | IFA: Fluorescence microscope at 200X or greater magnification | ELISA: Spectrophotometer: OD at 405/620 nm |
| Anti-EMA IgG cut-off | 2.5 Titer | ELISA: 20 EU/mL |
| Anti-EMA IgG semi-quantitative result | Endpoint Titer | ELISA: Negative: <20 EU/mL Indeterminate (Borderline): 20-25 EU/mL Positive: >25 EU/mL |
K. Standard/Guidance Document Referenced (if applicable):
None referenced.
L. Test Principle:
Using this indirect immunofluorescence method, patient serum is incubated on tissue sections to allow binding of antibodies to the substrate. Any antibodies not bound are removed by rinsing. Bound antibodies of the IgA or IgG class are detected by incubation of the substrate with fluorescein-labeled, anti-human immunoglobulin conjugates IgA or IgG respectively. Reactions are observed under a fluorescence microscope equipped with appropriate filters. The presence of EMA is demonstrated by an apple green fluorescence of the endomysium of smooth muscle bundles especially the inner layer. When positive, the titer (the reciprocal of the highest dilution giving a positive reaction) of the antibody is then determined by testing serial dilutions.
M. Performance Characteristics (if/when applicable):
1. Analytical performance:
a. Precision/Reproducibility:
The Anti-EMA IgA IFA intra-assay reproducibility was determined by testing three samples, i.e. low (2.5 titer), medium (20 titer), and high (1280 titer) ten times. All samples were within one titer difference. The IFA inter-assay reproducibility was determined by testing the same three samples in singlicate ten times in ten runs. All samples were within one titer difference.
The Anti-EMA IgG IFA intra-assay reproducibility was determined by testing three samples with high titer* (1280 – 20480) ten times. All samples were within one titer difference. The IFA inter-assay reproducibility was
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determined by testing the same three samples in singlicate ten times in ten runs. All samples were within one titer difference.
*Since anti-EMA IgG antibodies, when present, are generally in high titer, no patient samples close to the cut-off (usually difficult to obtain) were available for reproducibility studies.
b. Linearity/assay reportable range:
Not applicable
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
Not applicable.
d. Detection limit:
Not applicable.
e. Analytical specificity:
Interference by endogenous substances: The package insert states that grossly hemolyzed, lipemic, or microbially contaminated samples should not be used in these assays.
Cross-reactivity: Ten rheumatoid arthritis and ten systemic lupus erythematosus specimens were tested by both Anti-EMA IgA and IgG assays. All samples were negative.
f. Assay cut-off:
The cut-off value of 2.5 for EMA IgA was established using a population of 43 CD patients and 294 controls. Titers of CD patients ranged from 2.5 to >80. Titers of controls were all less than 2.5.
The cut-off value of 2.5 for EMA IgG was established using a population of 78 CD patients and 73 controls. Titers of CD patients ranged from 2.5 to 10240. Titers of controls were all less than 2.5.
2. Comparison studies:
a. Method comparison with predicate device:
Anti-EMA IgA - testing was performed on 215 samples which included 176 CD samples. The positive percent agreement was 98.9% (174/176); the negative percent agreement was 100% (39/39); and the overall agreement was 99.1% (213/215). Results are summarized below.
| | Anti-EMA IgA IFA (Primate Smooth Muscle) | | | |
| --- | --- | --- | --- | --- |
| | | Positive | Negative | Total |
| Anti-EMA-IgA IFA (Primate Distal Esophagus) | Positive | 174 | 0 | 174 |
| | Negative | 2 | 39 | 41 |
| | Total | 176 | 39 | 215 |
Positive percent Agreement: 98.9% (174/176)
Negative percent agreement: 100% (39/39)
Overall percent agreement: 99.1% (213/215)
The Anti-EMA IgG - testing was performed on 50 samples which included
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18 CD samples. The positive percent agreement was 83.3% (15/18); the negative percent agreement was 96.9% (31/32); and the overall agreement was 92% (46/50). Results are summarized below.
| | ImmuLisa Anti-tTG IgG (ELISA) | | | |
| --- | --- | --- | --- | --- |
| | | Positive | Negative | Total |
| Anti-EMA IgG IFA (Primate Distal Esophagus) | Positive | 15 | 1 | 16 |
| | Negative | 3 | 31 | 34 |
| | Total | 18 | 32 | 50 |
Positive percent Agreement: 83.3% (15/18)
Negative percent agreement: 96.9% (31/32)
Overall percent agreement: 92% (46/50)
b. Matrix comparison:
Not applicable.
3. Clinical studies:
a. Clinical Sensitivity and specificity:
The Anti-EMA IgA IFA clinical sensitivity and specificity study were evaluated on 60 clinically defined samples from patients with the following diagnosis: 30 Celiac Disease and 30 normal subjects. The Anti-EMA IgA IFA sensitivity was 100% (30/30) and the specificity 100% (30/30). Study results are summarized in the table below.
| | Diagnosis | | | |
| --- | --- | --- | --- | --- |
| | | Positive | Negative | Total |
| Anti-EMA IgA IFA (Primate Distal Esophagus) | Positive | 30 | 0 | 30 |
| | Negative | 0 | 30 | 30 |
| | Total | 30 | 30 | 60 |
Sensitivity: 100% (30/30)
Specificity: 100% (30/30)
The Anti-EMA IgG IFA clinical sensitivity and specificity study were evaluated on 24 clinically defined samples from patients with the following diagnosis: 14 Celiac Disease and 10 normal subjects. The Anti-EMA IgG IFA sensitivity was 100% (14/14) and the specificity 100% (10/10). Study results are summarized in the table below.
| | Diagnosis | | | |
| --- | --- | --- | --- | --- |
| | | Positive | Negative | Total |
| Anti-EMA IgG IFA (Primate Distal Esophagus) | Positive | 14 | 0 | 14 |
| | Negative | 0 | 10 | 10 |
| | Total | 14 | 10 | 24 |
Sensitivity: 100% (14/14)
Specificity: 100% (10/10)
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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.
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