(275 days)
An enzyme linked immunoassay (ELISA) for the qualitative or semi-quantitative detection of anti-centromere IgG antibodies in human serum as an aid in diagnosis of limited cutanteous systemic sclerosis / CREST in conjunction with other laboratory and clinical findings.
An enzyme linked immunoassay (EUSA) for the qualitative detection of anti-centromere IgG antibodies in human serum as an aid in diagnosis of limited cutaneous systemic sclerosis / CREST in conjunction with other laboratory and clinical findings.
This test is performed as a solid phase immunoassay. Microwells are coated with recombinant purified CENP-A centromere antigens. Controls, calibrators and patient sera are incubated in the antigen coated wells to allow specific antibodies present in the serum to bind to the centromere antibodies are detected by adding an enzyme labeled anti-human lgG conjugate. Specific enzyme substrate (TMB) is then added and the presence of antibodies is detected by a color change that is read by a spectrophotometer at 450 nm. Results are expressed in ELISA units per milliliter (EU/ml) and reported as positive or negative.
Here's an analysis of the provided text regarding the ImmuLisa™ Enhanced Centromere Antibody ELISA, focusing on acceptance criteria and supporting studies:
1. Table of Acceptance Criteria and Reported Device Performance
The document doesn't explicitly state formal "acceptance criteria" in a numerical target format (e.g., "sensitivity must be > X%"). Instead, the performance is demonstrated and presented for comparison to the predicate device and in clinical utility. Therefore, the reported performance metrics are presented in the table as the de facto "acceptance criteria" based on the study findings.
| Metric | Acceptance Criteria (Implied) | Reported Device Performance (ImmuLisa™) |
|---|---|---|
| Method Comparison (vs. Predicate INOVA QUANTA Lite™ Centromere ELISA) | ||
| Positive Percent Agreement (PPA) | High agreement with predicate | 93.2% (95% CI 82.7% - 97.8%) |
| Negative Percent Agreement (NPA) | High agreement with predicate | 95.4% (95% CI 92.5% - 97.3%) |
| Overall Agreement | High agreement with predicate | 95.1% (95% CI 92.4% - 96.9%) |
| Cross-Reactivity | Low false positives in other conditions | 2.7% (18/669 samples) |
| Precision | Low imprecision (CV%) across range | Total CV% range: 3.5% - 7.0% |
| Reproducibility (Qualitative) | High qualitative agreement | 62.5% for cutoff samples, 100% for ~20% above cutoff and other moderate positive samples. |
| Limit of Detection (LoD) | Clearly defined LoD | 3.9 EU/ml |
| Linearity and Recovery | Good linearity and recovery across assay range | Slope: 1.02-1.05, R²: 0.9842-0.9947, Recovery: 87%-116% |
| Hook Effect | No hook effect demonstrated | Not demonstrated up to 2531.3 EU/ml |
| Clinical Sensitivity (for IcSSc/CREST) | Demonstrate clinical utility | 53.2% (with 124 IcSSc/CREST samples) |
| Clinical Specificity (vs. autoimmune/infectious disease controls) | Demonstrate clinical utility | 95.5% (with 865 control samples) |
2. Sample Size Used for the Test Set and Data Provenance
- Method Comparison Test Set: 407 samples (59 positive, 348 negative by predicate).
- Cross-Reactivity Test Set: 669 specimens from individuals with various autoimmune or infectious diseases.
- Precision Test Set: 7 patient samples, run in duplicate, twice per day for 20 days (n=80 replicates per sample).
- Reproducibility Test Set: Samples ranging from ~20% below cutoff, ~20% above cutoff, and in the moderate positive range. Run in duplicate twice per day for 20 days.
- Limit of Detection Test Set: 60 replicates of blank, 10 replicates each of 6 low-level samples.
- Clinical Test Set: 124 IcSSc/CREST samples and 865 autoimmune and infectious disease controls.
Data Provenance: The document does not explicitly state the country of origin for the samples or whether they were retrospectively or prospectively collected. "Well-characterized lcSSc / CREST subjects and disease controls" are mentioned for the method comparison, suggesting pre-existing characterized samples. The clinical tests also use "Sets of clinical samples." Without further information, it's difficult to determine the specific provenance or collection method.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts
The document does not specify the number or qualifications of experts used to establish the ground truth for the test sets. For the method comparison, it refers to "well-characterized lcSSc / CREST subjects and disease controls," which implies clinical diagnosis, but the process of this characterization is not detailed. The "clinical tests" used IcSSc/CREST samples and controls, which would have had prior clinical diagnoses, but the adjudication of these diagnoses for the purpose of the study is not explained.
4. Adjudication Method for the Test Set
The document does not describe any specific adjudication method (e.g., 2+1, 3+1, none) for establishing the ground truth of the test set samples. The ground truth appears to be based on prior clinical diagnoses or characterization, but the details of how these were confirmed or adjudicated for the study are not provided.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done
No, a multi-reader multi-case (MRMC) comparative effectiveness study was not done. The device is an immunoassay (ELISA) for detecting antibodies, not an imaging or diagnostic algorithm that human readers would interact with.
6. If a Standalone (i.e., Algorithm Only Without Human-in-the-Loop Performance) Was Done
Yes, this is a standalone performance study. The ImmuLisa™ Enhanced Centromere Antibody ELISA is an in-vitro diagnostic device designed to provide results directly. Its performance (e.g., sensitivity, specificity, agreement) is assessed independently of direct human intervention in the result interpretation beyond standard laboratory procedures for running the assay and reading the spectrophotometer. The results are expressed quantitatively (EU/ml) and then interpreted qualitatively (positive/negative) based on a defined cutoff.
7. The Type of Ground Truth Used
The ground truth for the test sets (especially for method comparison and clinical performance) appears to be expert consensus / clinical diagnosis.
- For the Method Comparison, samples were "well-characterized lcSSc / CREST subjects and disease controls," implying pre-established clinical diagnoses.
- For Clinical Tests, "IcSSc/CREST samples" and "autoimmune and infectious disease controls" were used, again relying on existing clinical diagnoses as the reference standard.
8. The Sample Size for the Training Set
The document does not explicitly mention a "training set" in the context of device development or algorithm training. This is expected as the ImmuLisa™ is an ELISA kit, which relies on biochemical reactions and calibrated readouts rather than machine learning algorithms that typically require training data. The development of the assay itself would involve optimization and calibration, but this is distinct from "training a model."
9. How the Ground Truth for the Training Set Was Established
As no "training set" in the machine learning sense is described or implied for this ELISA kit, the question of how its ground truth was established is not applicable. The development of such diagnostic kits involves rigorous assay design, optimization, and validation against known standards and clinically characterized samples, but this is a different paradigm than algorithm training.
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Image /page/0/Picture/1 description: The image shows the seal of the U.S. Department of Health and Human Services. The seal features a stylized image of an eagle with three human profiles incorporated into its design. 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 Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
March 11, 2016
IMMCO Diagnostics Inc. Mr. Kevin Lawson Chief Regulatory Officer 60 Pineview Drive Buffalo, NY 14228
Re: K151559
Trade/Device Name: ImmuLisa™ Enhanced Centromere Antibody ELISA Regulation Number: 21 CFR §866.5100 Regulation Name: Antinuclear antibody immunological test system Regulatory Class: Class II Product Code: LJM, antinuclear antibody (enzyme-labeled), antigen, controls Dated: February 5, 2016 Received: February 5, 2016
Dear Mr. Lawson:
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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to 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 (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
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electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR 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 Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours,
Kelly Oliner - S
For
Leonthena R. Carrington, MBA, MS, MT(ASCP) Director
Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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| Form Approved: | OMB No. 0910-0120 |
|---|---|
| Expiration Date: | January 31, 2017 |
| See PRA Statement below. |
| 510(k) Number (if known) | K151559 |
|---|---|
| Device Name | ImmuLisa™ Enhanced Centromere Antibody ELISA |
| Indications for Use (Describe) | An enzyme linked immunoassay (ELISA) for the qualitative or semi-quantitative detection of anti-centromere IgG antibodies in human serum as an aid in diagnosis of limited cutanteous systemic sclerosis / CREST in conjunction with other laboratory and clinical findings. |
| Type of Use (Select one or both, as applicable) | Prescription Use (Part 21 CFR 801 Subpart D) Over-The-Counter Use (21 CFR 801 Subpart C) |
|---|---|
| ------------------------------------------------- | ---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- |
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Image /page/3/Picture/0 description: The image shows the logo for Immco Diagnostics. The logo features a cluster of blue and green dots on the left, followed by the company name in blue, with the word "DIAGNOSTICS" in green below. Underneath the company name is the text "A Trinity Biotech Company" in a smaller font.
510(k) Summary
| Submitter: | Immco Diagnostics, Inc. |
|---|---|
| Address: | 60 Pineview Dr., Buffalo, NY 14228 |
| Phone Number: | 716-691-0091 ext. 110 |
| Contact: | Kevin Lawson |
| Summary Prepared: | 3-6-2016 |
| Device Name: | ImmuLisa™ Enhanced Centromere Antibody ELISA |
| Common Name: | Centromere Antibody ELISA |
| Product Code: | LJM, Antinuclear Antibody (Enzyme-Labeled), Antigen, Control |
| Substantially Equivalent to: | INOVA QUANTA Lite™ Centromere ELISA |
General Description: Antinuclear antibodies (ANA) occur in sera of patients with various connective tissue disorders such as systemic lupus erythematosus (SLE), mixed connective tissue disease (MCTD), systemic sclerosis / scleroderma (SSc), polymyositis and Siögren's syndrome. These ANA are directed against nucleding DNA, nucleohistone and various extractable nuclear antigens (ENA) such as RNP, Sm, SS-A (Ro), SS-B (La), Centromere, Scl-70 (topoisomerase I) and Jo-1.
The anti-centromere antibody produces a discrete speckled pattern in HEp-2 immunofluorescence assays, and has been observed in 38-64% of patients with the limited cutaneous systemic sclerosis (IcSSc) or CREST syndrome variant of systemic sclerosis or scleroderma. Centromere antibodies bind the kinetochore of mitotic chromosomes and the pre-kinetochore of interphase cells. Immunoblotting analysis indicates that centromere antibodies recognize at least six epitopes on three chromosomal proteins: CENP-A (17-19kD), CENP-B (80kD), and CENP-C (140kD). CENP-A and CENP-B have been cloned and while CENP-B is the primary autoantigen, it appears that assays incorporating both antigens produce results with greater sensitivity and more comparable to immunofluorescence.
This test is performed as a solid phase immunoassay. Microwells are coated with recombinant purified CENP-A centromere antigens. Controls, calibrators and patient sera are incubated in the antigen coated wells to allow specific antibodies present in the serum to bind to the centromere antibodies are detected by adding an enzyme labeled anti-human lgG conjugate. Specific enzyme substrate (TMB) is then added and the presence of antibodies is detected by a color change that is read by a spectrophotometer at 450 nm. Results are expressed in ELISA units per milliliter (EU/ml) and reported as positive or negative.
Intended Use: An enzyme linked immunoassay (EUSA) for the qualitative detection of anti-centromere IgG antibodies in human serum as an aid in diagnosis of limited cutaneous systemic sclerosis / CREST in conjunction with other laboratory and clinical findings.
Similarities and Differences: Both kits use recombinant CENP-A and CENP-B coated on 96 well plates to detect IgG Centromere antibody with HRP anti-human IgG conjugate and TMB substrate. The IMMCO kit utilizes a 5 point calibrator curve with a borderline/indeterminate range of 20-25EU/ml; while the INOVA kit uses a single low positive sample as a cutoff of 20 units with no borderline range.
Non-clinical Tests:
Method Comparison: Both kits were tested with well-characterized lcSSc / CREST subjects and disease controls.
| INOVA Centromere Ab ELISA | ||||
|---|---|---|---|---|
| Positive | Negative | Total | ||
| IMMCOCentromere AbELISA | Positive | 55 | 16 | 71 |
| Negative | 4 | 332 | 336 | |
| Total | 59 | 348 | 407 |
60 Pineview Drive ● ● Buffalo, NY 14228-2120 ● ● USA Toll free (800) 537-8378 • tel. (716) 691-0091 • fax (716) 691-0466
www.immco.com
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Image /page/4/Picture/0 description: The image shows the logo for Immco Diagnostics, a Trinity Biotech Company. The logo features a cluster of blue and green circles on the left, followed by the company name "immco" in blue, with "DIAGNOSTICS" in green underneath. Below the company name is the text "A Trinity Biotech Company" in a smaller font size.
| Positive Percent Agreement: | 93.2% (95% Cl 82.7% - 97.8%) |
|---|---|
| Negative Percent Agreement: | 95.4% (95% CI 92.5% - 97.3%) |
| Overall Agreement: | 95.1% (95% CI 92.4% - 96.9%) |
Cross Reactivity:
A total of 669 potentially co-incident antibody positive specimens selected from individuals suffering from other autoimmune or infectious diseases were tested for centromere antibodies using the ImmuLisa™ assay. Indeterminate samples were considered positive.
| Condition | n | Centerome Ab Pos |
|---|---|---|
| Diffuse cutaneous scleroderma | 39 | 1 |
| Anti-phospholipid syndrome | 36 | 4 |
| Anti-phospholipid syndrome with SLE | 20 | 0 |
| Celiac Disease | 35 | 2 |
| Churg-Strauss syndrome | 27 | 0 |
| Crohn's disease | 25 | 0 |
| Granulomatosis with polyangitis | 27 | 0 |
| Graves' disease | 20 | 0 |
| Hashimoto's thyroiditis | 20 | 0 |
| Osteoarthritis | 20 | 0 |
| Polymyositis/Dermatomyositis | 30 | 1 |
| Rheumatoid Arthritis | 24 | 1 |
| Sjögren's Syndrome | 37 | 2 |
| Systemic Lupus Erythematosus | 92 | 1 |
| Thrombocytopenia | 15 | 2 |
| Ulcerative Colitis | 25 | 0 |
| Cytomegalovirus | 20 | 0 |
| Hepatitis C | 20 | 0 |
| Herpes simplex virus type 1 | 20 | 3 |
| Herpes simplex virus type 2 | 20 | 0 |
| Lyme disease | 20 | 0 |
| Mononucleosis | 20 | 1 |
| Rubella | 20 | 0 |
| Syphilis | 17 | 0 |
| Toxoplasmosis | 20 | 0 |
| Total | 669 | 18 (2.7%) |
Precision
Precision was tested with positive speciment the range of the assay. Seven patients were run in duplicate, twice per day for 20 days (n=80 replicates per sample). Assays were run by two operators on two different sets of equipment.
| S# | MeanEU/ml | TotalImprecision | Between days | BetweenOperator | Within run(Repeatability) | ||||
|---|---|---|---|---|---|---|---|---|---|
| SD | CV% | SD | CV% | SD | CV% | SD | CV% | ||
| 1 | 8.5 | 0.5 | 6.0 | 0.3 | 3.3 | 0.2 | 2.8 | 0.3 | 4.1 |
| 2 | 16.4 | 1.1 | 7.0 | 0.8 | 5.2 | 0.6 | 3.6 | 0.5 | 3.0 |
| 3 | 20.4 | 1.1 | 5.6 | 0.5 | 2.3 | 0.5 | 2.5 | 0.9 | 4.4 |
| 4 | 23.8 | 1.1 | 4.6 | 0.8 | 3.5 | 0.5 | 2.2 | 0.5 | 1.9 |
| 5 | 49.0 | 3.1 | 6.2 | 2.2 | 4.5 | 1.5 | 3.1 | 1.5 | 3.0 |
| 6 | 105.1 | 4.0 | 3.8 | 2.7 | 2.6 | 2.4 | 2.3 | 1.7 | 1.6 |
| 7 | 150.6 | 5.3 | 3.5 | 2.0 | 1.4 | 2.8 | 1.9 | 4.0 | 2.7 |
60 Pineview Drive ● USA Toll free (800) 537-8378 • tel. (716) 691-0091 • fax (716) 691-0466
www.immco.com
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Image /page/5/Picture/0 description: The image is a logo for Immco Diagnostics, a Trinity Biotech Company. The logo features a cluster of blue and green dots on the left side, followed by the word "immco" in blue, bold letters. Below "immco" is the word "DIAGNOSTICS" in green, and below that is the text "A Trinity Biotech Company" in a smaller font size.
Reproducibility
Qualitative reproducibility was tested with 80 runs of samples range, ~20% below cutoff, ~20% above cutoff, in the moderate positive range of the assay and near the qualitative analysis method. Samples were tested in duplicate twice per day for 20 days be two different operators / equipment sets. Assay results for the cutoff specimen produced 62.5% qualitative (positive) agreement. Assay results for the ~20% above cutoff speciment. All other speciment. All other specimens produced 100% qualitative agreement.
Limit of Detection
The limit of detection (LoD) was determined based on 60 replicates of the blank and 10 replicates each of 6 low-level (NHS) samples. LDD was determined to be 3.9 EU/ml.
Linearity and Recovery
Linearty and recovery were tested by diluting positive specimens through the assay range in equidistant dilutions and comparing actual vs. expected results. The linear range of the assay was determined be 3.9 (LoD) -160 EU/ml. Results are summarized below:
| TestRange(EU/ml) | Slope(95% CI) | Y-intercept(95% CI) | R2 | % recovery |
|---|---|---|---|---|
| 3.2 to 46.0 | 1.05 (0.97 to 1.13) | -0.67 (-2.88 to 1.53) | 0.9942 | 92% to 116% |
| 6.3 to 123.1 | 1.02 (0.95 to 1.10) | -0.18 (-5.64 to 5.29) | 0.9947 | 93% to 104% |
| 4.9 to 188.1 | 1.03 (0.95 to 1.11) | 1.42 (-7.15 to 9.98) | 0.9842 | 87% to 102% |
To assess hook effect, dilutions of high positive specimens with results above the 160 EU/ml measuring range were tested. Hook effect was not demonstrated in dilution samples with theoretical levels of 2531.3 EU/ml for testing within OD range of the microplate reader (~3.500).
Clinical Tests: Sets of clinical samples were tested on the IMMCO Centromere ELISA. By testing 124 IcSSc/CREST samples the sensitivity was determined to be 53.2%. By testing 865 autoimmune and infectious disease controls the specificity was determined to be 95.5%.
Kevin J. Lawson VP Regulatory Affairs
60 Pineview Drive Buffalo, NY 14228-2120 Toll free (800) 537-8378
§ 866.5100 Antinuclear antibody immunological test system.
(a)
Identification. An antinuclear antibody immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoimmune antibodies in serum, other body fluids, and tissues that react with cellular nuclear constituents (molecules present in the nucleus of a cell, such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis (a liver disease), rheumatoid arthritis, Sjögren's syndrome (arthritis with inflammation of the eye, eyelid, and salivary glands), and systemic sclerosis (chronic hardening and shrinking of many body tissues).(b)
Classification. Class II (performance standards).