(259 days)
INOVA QUANTA Lite™ ß2 GPI IgA, IgG, IgM and Screen ELISA
Not Found
No
The device description details a standard ELISA assay, which is a biochemical test. There is no mention of AI or ML in the device description, intended use, or performance studies. The analysis of results is based on spectrophotometry and conversion to ELISA units, not algorithmic interpretation.
No
This device is an in vitro diagnostic test for detecting antibodies related to autoimmune disorders, not a device used for therapy.
Yes
The 'Intended Use / Indications for Use' section explicitly states that the device is used "to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE)".
No
The device description clearly outlines a physical immunoassay (ELISA) process involving reagents, microwells, incubation, washing, and spectrophotometric reading. This is a hardware-based in vitro diagnostic test, not a software-only device.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use clearly states that the device is an "Enzyme-linked immunosorbent assay (ELISA) for the qualitative or semi-quantitative detection of β2-GPI IgA, IgG, and/or IgM antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings." This describes a test performed on a biological sample (human serum) outside of the body to provide information for diagnosis.
- Device Description: The description details a laboratory test method (ELISA) performed on serum samples.
- Performance Studies: The document includes descriptions of performance studies conducted on human serum samples (Method Comparison, Cross Reactivity, Clinical Tests) to evaluate the device's ability to detect the target antibodies and its diagnostic performance.
All of these characteristics align with the definition of an In Vitro Diagnostic device.
N/A
Intended Use / Indications for Use
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Enzyme-linked immunosorbent assay (ELISA) for the qualitative or semi-quantitative detection of β2-GPI IgA antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings.
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Enzyme-linked immunosorbent assay (ELISA) for the qualitative or semi-quantitative detection of β2-GPI IgG antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings.
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Enzyme-linked immunosorbent assay (ELISA) for the qualitative or semi-quantitative detection of β2-GPI IgM antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings.
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Enzyme-linked immunosorbent assay (ELISA) for the qualitative detection of β2-GPI IgA, IgG and IgM antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings.
Product codes (comma separated list FDA assigned to the subject device)
MSV
Device Description
Antiphospholipid are a heterogeneous group of autoantibodies against negatively charged phospholipids. They are detected primarily by the anti-cardiolipin antibody (ACA) test, the biological false positive test for syphilis or the lupus anticoagulant test. These three tests detect related, but not necessarily identical antibodies. Thus, more than one of these tests may be necessary to identify antiphospholipid antibodies. Of the various tests for the detection of antibodies, the anti-cardiolipin antibody test performed by ELISA is the most sensitive." The presence of anti-cardiolipin antibodies helps to identify indents at risk of venous and/or arterial thrombosis often accompanied by thrombocytopenia, a syndrome referred to as antiphospholipid syndrome." 12tt most commonly occurs in patients with systemic lupus erythematosus (SLE) or lupus-like diseases where the criteria for SLE are not fulfilled." High levels of anti-cardiolipin antibodies occur in thrombocytopenia and several other disorders. 43 Anti-cardiolipin antibodies are routinely detected by ELISA, using cardiolipin as the antigen. Recent observations have indicated that 50kD serum factor is necessary for ACA to bind to cardiolipin. This co-factor was later identified as beta 2 glycoprotein 1 (82GP1) or synonymously apolipoprotein H. 488 Though the function of B2GP1 remains unclear, it is certain that the presence of B2GP1 facilitates the binding of the ACA to cardiolipin antibodies, especially at low levels are found in a variety of clinical disorders unrelated to anti-phospholipid syndrome. ACA present in sera of patients with SLE and other are immunologically distinct from sera of patients with syphilis. The use of B2GP1 protein as a replacement for cardiolipin antigen on the solid matrix helps to make this distinction. Furthermore, anti-B2GP1 antibodies are very specific for anti-phospholipid syndrome The test is performed as a solid phase immunoassay (ELISA) in B2GP1 coated microwells. Controls, Calibrators and patient serum samples are incubated in the antigen coated microwells to allow antibodies present in the serum to bind. Unbound antibody and other serum proteins are removed by washing the microwells are detected by adding an enzyme labeled anti-human lgA, lgG, lgM or lgA/lgG/lgM conjugate to the microwells. These enzyme conjugated antibodies bind specifically to the human immunoglobulin of the apropriate class. Unbound enzyme-labeled conjugate is removed by washing. Specific enzyme substrate (TMB) is then added to the presence of antibodies is detected by a color change produced by the conversion of the TMB substrate. The reaction is stopped and the intensity of color change, which is proportional to the concentration of antibody, is read by a spectrophotometer at 450 nm. Results are expressed in ELISA units per milliliter (EU/ml).
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
human serum
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Not Found
Description of the training set, sample size, data source, and annotation protocol
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Description of the test set, sample size, data source, and annotation protocol
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Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Method Comparison: Both kits were tested with APS subjects and disease controls.
IMMCO IgA Method Comparison Study 2x2 Table:
Total sample size: 328
Positive % Agreement: 74.8% (95% CI 65.7% - 82.2%)
Negative % Agreement: 99.1% (95% CI 96.3% - 99.8%)
Overall % Agree: 90.5% (95% CI 86.7% - 93.4%)
IMMCO IgG Method Comparison Study 2x2 Table:
Total sample size: 338
Positive % Agreement: 83.1% (95% CI 72.5%-90.4%)
Negative % Agreement: 92.0% (95% CI 87.8%-94.8%)
Overall % Agree: 89.9% (95% CI 86.2%-92.8%)
IMMCO IgM Method Comparison Study 2x2 Table:
Total sample size: 355
Positive % Agreement: 91.7% (95% CI 84.8% - 95.7%)
Negative % Agreement: 99.1% (95% CI 96.7% - 99.9%)
Overall % Agree: 96.6% (95% CI 94.3% - 98.4%)
IMMCO IgA/G/M Method Comparison Study 2x2 Table:
Total sample size: 331
Positive % Agreement: 80.2% (95% CI 73.4%-85.7%)
Negative % Agreement: 96.8% (95% CI 92.2%-98.8%)
Overall % Agree: 87.9% (95% CI 83.8%-91.1%)
Cross Reactivity: Potentially co-incident antibody positive specimens selected from individuals suffering from other autoimmune or infectious diseases were tested for B2GP1 antibodies using the ImmuLisa™ assay. Indeterminate samples were considered positive. Tables provided for IgA, IgG, IgM, and IgA/IgG/IgM with various conditions and n Pos/% Pos.
Interference: Interference was studied by mixing sera with known B2GP1 levels for each assay with potentially interfering serum samples and studying deviation from expected results. No significant interference was demonstrated for the levels indicated: Hemoglobin (2 g/L), Bilirubin (342 umo/L), Rheumatoid Factor (100 EU/ml), Triglycerides (37 mmo/4), Heparin (3000 U/L), Acetylsalicylic Acid (3.62 mmol/L), Warfarin (32.5 µmo/L), Hydroxychloroquine (148 µmo//L), Rituximal (75 μg/ml), and Atorvastatin (600 μg ΕΩ/L).
Precision: Precision was tested with positive specimens selected throughout the range of the assay. Seven patients were run in 13 assays with 6 replicates of each sample over a period of 4 weeks. An additional assay of 12 replicates was run to determine repeatability (n=90 replicates per sample). Assays were run by two operators on two different sets of equipment.
Results summarized in a table showing Mean EU/ml, Total Imprecision, Between days, Between Operator, and Within run (Repeatability) with values and CV%.
Reproducibility: Qualitative reproducibility was tested with 90 replicates of samples in the negative range, ~20% below cutoff, in the moderate positive range of the assay and near the qualitative analysis method. Samples were tested by two different operators / equipment sets. The cutoff samples produced 76.7% (positive) and 61.1% (negative) qualitative agreement for Ize, JgG and IgM, respectively. Results for the 10% specimen produced 96.7% (negative) qualitative agreement. Assay results for all other specimens produced 100% qualitative agreement. For the 120% below cutoff specimen produced 97.8% qualitative agreement (negative). The cutoff sample produced 50% negative qualitative agreement. Results for all other specimens produced 100% qualitative agreement.
Limit of Detection: The limits of detection (LoD) for these assays were determined to be 3.6 EU/ml for lgG , 2.3 EU/ml for lgM, and 2.7 EU/ml for IgA/IgG/IgM based on 60 replicates of the blank and 10 replicates each of 5 low-level (normal blood donor) samples.
Linearity and Recovery: For the individual IgA, IgG and IgM kits, linearity and recovery were tested by diluting positive sssay range in equidistant dilutions and comparing actual results. The linear range of the lgA assay was determined to be 3.6–160 EU/ml. The linear range of the lgG assay was determined to be 3.1-160 EU/ml. The linear range of the IgM assay was determined to be 2.3-160 EU/ml.
Results summarized in tables for B2GP1 IgA, B2GP1 IgG, and B2GP1 IgM showing Test Range (EU/ml), Slope (95% CI), Y-intercept (95% CI), R2, and % recovery (obtained/expected).
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 as high as 228.6 EU/ml for IgG, and 605.5 EU/ml for IgM testing within OD range of the microplate reader (~3.50D).
Clinical Tests: Sets of clinical samples were tested to assess sensitivity and specificity for APS, APS with SLE and SLE. Results are summarized below.
Sensitivity/specificity exclude healthy human blood on the reference laboratory testing with a possible diagnosis of APS or SLE. Borderline results are considered positive.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Clinical Performance:
For APS:
B2GP1 IgA: Sensitivity 38.3 (30.0-47.3), Specificity 92.0 (89.3-94.1)
B2GP1 IgG: Sensitivity 58.8 (48.3-68.5), Specificity 93.8 (91.3-95.7)
B2GP1 IgM: Sensitivity 64.1 (55.7-71.8), Specificity 96.1 (93.9-97.5)
B2GP1 IgA/IgG/IgM: Sensitivity 84.8% (76.1-90.8), Specificity 91.9% (89.1-94.0)
For APS with SLE:
B2GP1 IgA: Sensitivity 19.4 (12.5-28.6), Specificity 92.0 (89.3-94.1)
B2GP1 IgG: Sensitivity 57.7 (48.5-66.5), Specificity 93.8 (91.3-95.7)
B2GP1 IgM: Sensitivity 46.5 (36.5-56.7), Specificity 96.1 (93.9-97.5)
B2GP1 IgA/IgG/IgM: Sensitivity 66.0% (55.8-75.0), Specificity 91.9% (89.1-94.0)
For SLE:
B2GP1 IgA: Sensitivity 10.9 (4.9-21.8), Specificity 92.0 (89.3-94.1)
B2GP1 IgG: Sensitivity 26.5 (18.4-36.6), Specificity 93.8 (91.3-95.7)
B2GP1 IgM: Sensitivity 18.8 (12.0-28.1), Specificity 96.1 (93.9-97.5)
B2GP1 IgA/IgG/IgM: Sensitivity 24.6% (14.8-37.6), Specificity 91.9% (89.1-94.0)
Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
INOVA QUANTA Lite™ ß2 GPI IgA, IgG, IgM and Screen ELISA
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 866.5660 Multiple autoantibodies immunological test system.
(a)
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).(b)
Classification. Class II (performance standards).
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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is a stylized image of three human profiles facing to the right, with flowing lines representing hair or movement.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
June 19, 2017
IMMCO DIAGNOSTICS, INC. Kevin Lawson Vice President, Regulatory Affairs 9870 Hollingson Road Clarence, NY 14031
Re: K162788
Trade/Device Name: ImmuLisa Enhanced B2GP1 IgA Antibody ELISA, ImmuLisa Enhanced B2GP1 IgG Antibody ELISA, ImmuLisa Enhanced B2GP1 IgM Antibody ELISA, ImmuLisa Enhanced B2GP1 IgA/IgG/IgM Antibody ELISA Regulation Number: 21 CFR 866.5660 Regulation Name: Multiple autoantibodies immunological test system Regulatory Class: II Product Code: MSV Dated: May 30, 2017 Received: May 31, 2017
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
1
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 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 regulations (21 CFR Parts 801 and 809). 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/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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 Carrington, MBA, MS, MT (ASCP) Division 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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Indications for Use
510(k) Number (if known) K162788
Device Name
- ImmuLisa Enhanced™ B2GP1 IgA Antibody ELISA. 2. ImmuLisa Enhanced™ B2GP1 IgG Antibody ELISA. 3. ImmuLisa Enhanced™ B2GP1 IgM Antibody ELISA. 4. ImmuLisa Enhanced™ B2GP1 IgA/IgG/IgM ELISA.
Indications for Use (Describe)
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Enzyme-linked immunosorbent assay (ELISA) for the qualitative or semi-quantitative detection of β2-GPI IgA antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings.
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Enzyme-linked immunosorbent assay (ELISA) for the qualitative or semi-quantitative detection of β2-GPI IgG antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings.
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Enzyme-linked immunosorbent assay (ELISA) for the qualitative or semi-quantitative detection of β2-GPI IgM antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings.
-
Enzyme-linked immunosorbent assay (ELISA) for the qualitative detection of β2-GPI IgA, IgG and IgM antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests 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, a Trinity Biotech Company. The logo features a cluster of blue and green dots on the left, followed by the company name "immco" in blue, and the word "DIAGNOSTICS" in green underneath. Below that, the text "A Trinity Biotech Company" is written 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: | 6-14-2017 |
| Device Name: | ImmuLisa Enhanced™ B2GP1 IgA Antibody ELISA |
| ImmuLisa Enhanced™ B2GP1 IgG Antibody ELISA | |
| ImmuLisa Enhanced™ B2GP1 IgM Antibody ELISA | |
| ImmuLisa Enhanced™ B2GP1 IgA/IgG/IgM Antibody ELISA | |
| Common Name: | B2GP1 Antibody ELISA |
| Product Code: | MSV, system,test,antibodies,b2 - glycoprotein i (b2 - gpi) |
| Substantially Equivalent to: | INOVA QUANTA Lite™ ß2 GPI IgA, IgG, IgM and Screen ELISA |
General Description: Antiphospholipid are a heterogeneous group of autoantibodies against negatively charged phospholipids. They are detected primarily by the anti-cardiolipin antibody (ACA) test, the biological false positive test for syphilis or the lupus anticoagulant test. These three tests detect related, but not necessarily identical antibodies. Thus, more than one of these tests may be necessary to identify antiphospholipid antibodies.
Of the various tests for the detection of antibodies, the anti-cardiolipin antibody test performed by ELISA is the most sensitive." The presence of anti-cardiolipin antibodies helps to idents at risk of venous and/or arterial thrombosis often accompanied by thrombocytopenia, a syndrome referred to as antiphospholipid syndrome." 12 tt most commonly occurs in patients with systemic lupus erythematosus (SLE) or lupus-like diseases where the criteria for SLE are not fulfilled." High levels of anti-cardiolipin antibodies occur in thrombocytopenia and several other disorders. 43 Anti-cardiolipin antibodies are routinely detected by ELISA, using cardiolipin as the antigen. Recent observations have indicated that 50kD serum factor is necessary for ACA to bind to cardiolipin. This co-factor was later identified as beta 2 glycoprotein 1 (82GP1) or synonymously apolipoprotein H. 488 Though the function of B2GP1 remains unclear, it is certain that the presence of B2GP1 facilitates the binding of the ACA to cardiolipin antibodies, especially at low levels are found in a variety of clinical disorders unrelated to anti-phospholipid syndrome. ACA present in sera of patients with SLE and other are immunologically distinct from sera of patients with syphilis. The use of B2GP1 protein as a replacement for cardiolipin antigen on the solid matrix helps to make this distinction. Furthermore, anti-B2GP1 antibodies are very specific for anti-phospholipid syndrome
The test is performed as a solid phase immunoassay (ELISA) in B2GP1 coated microwells. Controls, Calibrators and patient serum samples are incubated in the antigen coated microwells to allow antibodies present in the serum to bind. Unbound antibody and other serum proteins are removed by washing the microwells are detected by adding an enzyme labeled anti-human lgA, lgG, lgM or lgA/lgG/lgM conjugate to the microwells. These enzyme conjugated antibodies bind specifically to the human immunoglobulin of the apropriate class. Unbound enzyme-labeled conjugate is removed by washing. Specific enzyme substrate (TMB) is then added to the presence of antibodies is detected by a color change produced by the conversion of the TMB substrate. The reaction is stopped and the intensity of color change, which is proportional to the concentration of antibody, is read by a spectrophotometer at 450 nm. Results are expressed in ELISA units per milliliter (EU/ml).
Intended Use: Immulisa Enhanced™ B2GP1 IgA Antibody ELISA: Enzyme-linked immunosorbent assay (EUSA) for the qualitative or semi-quantitative detection of β2-GPI IgA antibodies in human serum to aitoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings.
ImmuLisa Enhanced™ 82GP1 IgA/IgG/IGM Antibody ELISA: Enzyme-linked immunosorbent assay (ELISA) for the qualitative detection of B2-GPI IgA, IgG and IgM antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with
60 Pineview Drive ● Toll free (800) 537-8378 ● tel. (716) 691-0091 ● fax (716) 691-0466
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Image /page/4/Picture/0 description: The image shows the logo for Immco Diagnostics. The logo features a cluster of blue and green circles on the left, followed by the word "immco" in blue, with a small circle above the "o". Below "immco" is the word "DIAGNOSTICS" in green. Underneath that, it says "A Trinity Biotech Company" in a smaller font.
antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings.
Immulisa Enhanced™ B2GP1 IgM Antibody ELISA: Enzyme-linked immunosorbent assay (ELISA) for the qualitative or semiquantitative detection of $2-GPI IgM antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings.
Immulisa Enhanced™ B2GP1 IgG Antibody ELISA: Enzyme-linked immunosorbent assay (ELISA) for the qualitative or semiquantitative detection of ß2-GPI IgG antibodies in human serum to aid in diagnosis of autoimmune thrombotic disorders associated with antiphospholipid syndrome (APS) and APS with systemic lupus erythematosus (SLE) in conjunction with other laboratory tests and clinical findings.
Similarities and Differences: Both sets of kits detect antibodies to beta 2 glycoprotein 1antigen coated on 96 well plates to detect with HRP anti-human IgG conjugate and TMB substrate. Both sets of IgA, JgG and IgM kits utilize a 5 point calibrator curve, however, Immco uses a borderline/indeterminate range of 20-25EU/ml, while INOVA does not use a borderline/indeterminate range. The lmmco lgA/gG/IGM kit uses a single calibrator set at 30EU/ml, has a cutoff of 20 units with no borderline range and incorporates a combined IgA/IgG/IgM conjugate.
Non-clinical Tests:
Method Comparison: Both kits were tested with APS subjects and disease controls.
IMMCO IgA Method Comparison Study 2x2 Table
| borderline considered positive | Other B2GP1 IgA ELISA | ||||
|---|---|---|---|---|---|
| Pos | Neg | Total | |||
| IMMCO | Pos | 86 | 2 | 88 | |
| B2GP1 IgA | Neg | 29 | 211 | 240 | |
| ELISA | Total | 115 | 213 | 328 | |
| Pos % Agreement | 74.8% | (95% CI 65.7% - 82.2%) | |||
| Neg % Agreement | 99.1% | (95% CI 96.3% - 99.8%) | |||
| Overall % Agree | 90.5% | (95% CI 86.7% - 93.4%) |
IMMCO IgG Method Comparison Study 2x2 Table
borderline considered positive
| Other B2GP1 IgG ELISA | ||||
|---|---|---|---|---|
| Pos | Neg | Total | ||
| IMMCO | Pos | 64 | 21 | 85 |
| B2GP1 IgG | ||||
| ELISA | Neg | 13 | 240 | 253 |
| Total | 77 | 261 | 338 | |
| Pos % Agreement | 83.1% | (95% CI 72.5%-90.4%) | ||
| Neg % Agreement | 92.0% | (95% CI 87.8%-94.8%) | ||
| Overall % Agree | 89.9% | (95% CI 86.2%-92.8%) |
60 Pineview Drive ● Buffalo, NY 14228-2120 USA Toll free (800) 537-8378 ● tel. (716) 691-0091 ● fax (716) 691-0466
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Image /page/5/Picture/0 description: The image shows the logo for Immco Diagnostics, a Trinity Biotech company. 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. The phrase "A Trinity Biotech Company" is written in a smaller font size below the company name.
IMMCO IgM Method Comparison Study 2x2 Table
borderline considered positive
| Other B2GP1 IgM ELISA | ||||
|---|---|---|---|---|
| Pos | Neg | Total | ||
| IMMCO | Pos | 110 | 2 | 112 |
| B2GP1 IgM | Neg | 10 | 233 | 243 |
| ELISA | Total | 120 | 235 | 355 |
| Pos % Agreement | 91.7% | (95% CI 84.8% - 95.7%) | ||
| Neg % Agreement | 99.1% | (95% CI 96.7% - 99.9%) | ||
| Overall % Agree | 96.6% | (95% CI 94.3% - 98.4%) |
IMMCO IgA/G/M Method Comparison Study 2x2 Table
| Other IgA/G/M specific isotype ELISA | |||||
|---|---|---|---|---|---|
| Any Pos | All Neg | Total | |||
| IMMCO | Pos | 142 | 5 | 147 | |
| B2GP1 | Neg | 35 | 149 | 184 | |
| A/G/M ELISA | Total | 177 | 154 | 331 | |
| Pos % Agreement | 80.2% | (95% CI 73.4%-85.7%) | |||
| Neg % Agreement | 96.8% | (95% CI 92.2%-98.8%) | |||
| Overall % Agree | 87.9% | (95% CI 83.8%-91.1%) |
Cross Reactivity:
Potentially co-incident antibody positive specimens selected from individuals suffering from other autoimmune or infectious diseases were tested for B2GP1 antibodies using the ImmuLisa™ assay. Indeterminate samples were considered positive.
| B2GP1 Abs | IgA | IgG | IgM | |||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Condition | n | n Pos | % Pos | n | n Pos | % Pos | n | n Pos | % Pos | |
| Syphilis | 40 | 7 | 17.5% | 40 | 3 | 7.5% | 40 | 5 | 12.5% | |
| HCV | 32 | 0 | 0.0% | 32 | 3 | 9.4% | 32 | 1 | 3.1% | |
| Celiac | 16 | 0 | 0.0% | 0 | 0 | 0.0% | 0 | 0 | 0.0% | |
| Polymyositis | 17 | 3 | 17.6% | 17 | 0 | 0.0% | 17 | 1 | 5.9% | |
| Dermatomyositis | 12 | 2 | 16.7% | 12 | 0 | 0.0% | 12 | 1 | 8.3% | |
| RA | 46 | 6 | 13.0% | 60 | 4 | 6.7% | 60 | 0 | 0.0% | |
| Systemic Sclerosis | 31 | 3 | 9.7% | 23 | 3 | 13.0% | 23 | 1 | 4.3% | |
| Sjögren's | 37 | 2 | 5.4% | 37 | 5 | 13.5% | 37 | 2 | 5.4% | |
| Stroke | 15 | 1 | 6.7% | 15 | 0 | 0.0% | 15 | 1 | 6.7% | |
| Epilepsy | 14 | 1 | 7.1% | 14 | 0 | 0.0% | 14 | 1 | 7.1% | |
| Arterial Thrombosis | 15 | 1 | 6.7% | 15 | 2 | 13.3% | 15 | 1 | 6.7% | |
| Venous Thrombosis | 15 | 0 | 0.0% | 15 | 0 | 0.0% | 15 | 0 | 0.0% | |
| Myocardial Infarction | 20 | 3 | 15.0% | 20 | 0 | 0.0% | 20 | 1 | 5.0% | |
| Acute Coronary Syndrome | 10 | 1 | 10.0% | 10 | 0 | 0.0% | 10 | 0 | 0.0% | |
| Pre-eclampsia | 20 | 0 | 0.0% | 20 | 3 | 15.0% | 20 | 0 | 0.0% | |
| SNHL | 0 | 0 | 0.0% | 10 | 0 | 0.0% | 0 | 0 | 0.0% | |
| Hashimoto's | 7 | 0 | 0.0% | 20 | 0 | 0.0% | 20 | 0 | 0.0% | |
| Graves | 25 | 3 | 12.0% | 18 | 1 | 5.6% | 18 | 0 | 0.0% | |
| Wegener's | 45 | 4 | 8.9% | 34 | 0 | 0.0% | 34 | 1 | 2.9% |
60 Pineview Drive ● Buffalo, NY 14228-2120 ● USA Toll free (800) 537-8378 ● tel. (716) 691-0091 ●
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Image /page/6/Picture/0 description: The image shows the logo for Immco Diagnostics. The logo features a cluster of blue and green circles on the left, followed by the word "immco" in blue, with a small circle above the "o". Below "immco" is the word "DIAGNOSTICS" in green, and below that is the text "A Trinity Biotech Company".
| Churg-Strauss | 23 | 3 | 13.0% | 23 | 1 | 4.3% | 23 | 0 |
|---|---|---|---|---|---|---|---|---|
| Crohn's | 20 | 0 | 0.0% | 20 | 1 | 5.0% | 20 | 0 |
| Ulcerative Colitis | 20 | 1 | 5.0% | 20 | 2 | 10.0% | 20 | 2 |
| Toxoplasma | 10 | 0 | 0.0% | 10 | 1 | 10.0% | 10 | 1 |
| CMV | 10 | 0 | 0.0% | 10 | 1 | 10.0% | 10 | 1 |
| Rubella | 10 | 0 | 0.0% | 10 | 0 | 0.0% | 10 | 0 |
| B2GP1 Abs | IgA/IgG/IgM | |||||||
| Condition | n | n Pos | % Pos | |||||
| Syphilis | 37 | 8 | 21.6% | |||||
| HCV | 32 | 2 | 6.3% | |||||
| Celiac | 22 | 1 | 4.5% | |||||
| Polymyositis | 17 | 2 | 11.8% | |||||
| Dermatomyositis | 12 | 0 | 0.0% | |||||
| RA | 45 | 3 | 6.7% | |||||
| Systemic Sclerosis | 23 | 2 | 8.7% | |||||
| Sjögren's | 37 | 2 | 5.4% | |||||
| Stroke | 15 | 1 | 6.7% | |||||
| Epilepsy | 14 | 1 | 7.1% | |||||
| Arterial Thrombosis | 15 | 3 | 20.0% | |||||
| Venous Thrombosis | 15 | 1 | 6.7% | |||||
| Myocardial Infarction | 20 | 4 | 20.0% | |||||
| Acute Coronary Syndrome | 10 | 1 | 10.0% | |||||
| Pre-eclampsia | 20 | 0 | 0.0% | |||||
| SNHL | 7 | 0 | 0.0% | |||||
| Hashimoto's | 7 | 0 | 0.0% | |||||
| Graves | 20 | 1 | 5.0% | |||||
| Wegener's | 42 | 1 | 2.4% | |||||
| Churg-Strauss | 23 | 2 | 8.7% | |||||
| Crohn's | 20 | 1 | 5.0% | |||||
| Ulcerative Colitis | 20 | 2 | 10.0% | |||||
| Toxoplasma | 10 | 1 | 10.0% | |||||
| CMV | 10 | 2 | 20.0% |
Interference
| 10
0
0.0%
Rubella
Interference was studied by mixing sera with known B2GP1 levels for each assay with potentially interfering serum samples and studying deviation from expected results. No significant interference was demonstrated for the levels indicated: Hemoglobin (2 g/L), Bilirubin (342 umo/L), Rheumatoid Factor (100 EU/ml), Triglycerides (37 mmo/4), Heparin (3000 U/L), Acetylsalicylic Acid (3.62 mmol/L), Warfarin (32.5 µmo/L), Hydroxychloroquine (148 µmo//L), Rituximal (75 μg/ml), and Atorvastatin (600 μg ΕΩ/L).
0.0% 0.0% 10.0% 10.0% 10.0% 0.0%
Precision
Precision was tested with positive specimens selected throughout the range of the assay. Seven patients were run in 13 assays with 6 replicates of each sample over a period of 4 weeks. An additional assay of 12 replicates was run to determine repeatability (n=90 replicates per sample). Assays were run by two operators on two different sets of equipment.
60 Pineview Drive USA Toll free (800) 537-8378 ● tel. (716) 691-0091 ● fax (716) 691-0466
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| S# | Mean
EU/ml | Total
Imprecision | | Between days | | Between
Operator | | Within run
(Repeatability) | |
|----|---------------|----------------------|-----|--------------|-----|---------------------|-----|-------------------------------|-----|
| 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 |
Reproducibility
Qualitative reproducibility was tested with 90 replicates of samples in the negative range, ~20% below cutoff, in the moderate positive range of the assay and near the qualitative analysis method. Samples were tested by two different operators / equipment sets. The cutoff samples produced 76.7% (positive) and 61.1% (negative) qualitative agreement for Ize, JgG and IgM, respectively. Results for the 10% specimen produced 96.7% (negative) qualitative agreement. Assay results for all other specimens produced 100% qualitative agreement. For the 120% below cutoff specimen produced 97.8% qualitative agreement (negative). The cutoff sample produced 50% negative qualitative agreement. Results for all other specimens produced 100% qualitative agreement.
Limit of Detection
The limits of detection (LoD) for these assays were determined to be 3.6 EU/ml for lgG , 2.3 EU/ml for lgM, and 2.7 EU/ml for IgA/IgG/IgM based on 60 replicates of the blank and 10 replicates each of 5 low-level (normal blood donor) samples. .
Linearity and Recovery
For the individual IgA, IgG and IgM kits, linearity and recovery were tested by diluting positive sssay range in equidistant dilutions and comparing actual results. The linear range of the lgA assay was determined to be 3.6–160 EU/ml. The linear range of the lgG assay was determined to be 3.1-160 EU/ml. The linear range of the IgM assay was determined to be 2.3-160 EU/ml. Results are summarized below:
| Test Range
(EU/ml) | Slope
(95% CI) | Y-intercept
(95% CI) | R2 | % recovery
(obtained/expected) |
|-----------------------|---------------------|-------------------------|--------|-----------------------------------|
| B2GP1 IgA | | | | |
| 2.3 to 17.8 | 1.06 (0.89 to 1.21) | -0.59 (-2.4 to 1.2) | 0.9767 | 88% to 112% |
| 11.0 to 63.9 | 1.05 (0.97 to 1.14) | -0.77 (-4.2 to 2.6) | 0.9934 | 93% to 105% |
| 59.0 to 168.3 | 1.08 (0.94 to 1.22) | -0.68 (-16.7 to 15.4) | 0.9915 | 90% to 100% |
| B2GP1 IgG | | | | |
| 2.1 to 41.4 | 1.04 (0.89 to 1.19) | 0.2 (-1.8 to 4.9)) | 0.9807 | 82% to 100% |
| 4.1 to 140.9 | 1.03 (0.93 to 1.13) | 4.13 (-3.9 to 12.1) | 0.9905 | 86% to 100% |
| 3.3 to 198.1 | 0.99 (0.91 to 1.07) | 7.25 (-1.8 to 16.3) | 0.9936 | 82% to 100% |
| B2GP1 IgM | | | | |
| 1.7 to 63.4 | 1.00 (0.95 to 1.05) | 1.8 (-0.1 to 3.6) | 0.9974 | 81% to 100% |
| 5.4 to 87.5 | 1.02 (0.93 to 1.10) | 2.3 (-1.9 to 6.4) | 0.9936 | 89% to 100% |
| 54.7 to 173.5 | 0.96 (0.82 to 1.09) | 3.1 (-13.1 to 19.4) | 0.9963 | 86% to 110% |
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 as high as 228.6 EU/ml for IgG, and 605.5 EU/ml for IgM testing within OD range of the microplate reader (~3.50D).
60 Pineview Drive Buffalo, NY 14228-2120 Toll free (800) 537-8378 • tel. (716) 691-0091 • fax (716) 691-0466
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Image /page/8/Picture/0 description: The image shows the logo for Immco Diagnostics, a Trinity Biotech Company. The logo features a cluster of blue and green dots to the left of the word "immco" in blue, with "DIAGNOSTICS" in green below it. Underneath that is the text "A Trinity Biotech Company" in a smaller font.
Clinical Tests: Sets of clinical samples were tested to assess sensitivity and specificity for APS, APS with SLE and SLE. Results are summarized below.
| Disease | B2GP1 IgA % (95%CI) | B2GP1 IgG % (95%CI) | B2GP1 IgM% (95%CI) | B2GP1 IgA/IgG/IgM% (95%CI) | ||||
|---|---|---|---|---|---|---|---|---|
| Sensitivity | Specificity | Sensitivity | Specificity | Sensitivity | Specificity | Sensitivity | Specificity | |
| APS | 38.3 (30.0-47.3) | 92.0 (89.3-94.1) | 58.8 (48.3-68.5) | 93.8 (91.3-95.7) | 64.1 (55.7-71.8) | 96.1 (93.9-97.5) | 84.8% (76.1-90.8) | 91.9% (89.1-94.0) |
| APS with SLE | 19.4 (12.5-28.6) | 92.0 (89.3-94.1) | 57.7 (48.5-66.5) | 93.8 (91.3-95.7) | 46.5 (36.5-56.7) | 96.1 (93.9-97.5) | 66.0% (55.8-75.0) | 91.9% (89.1-94.0) |
| SLE | 10.9 (4.9-21.8) | 92.0 (89.3-94.1) | 26.5 (18.4-36.6) | 93.8 (91.3-95.7) | 18.8 (12.0-28.1) | 96.1 (93.9-97.5) | 24.6% (14.8-37.6) | 91.9% (89.1-94.0) |
- Sensitivity/specificity exclude healthy human blood on the reference laboratory testing with a possible diagnosis of APS or SLE. Borderline results are considered positive.
Kulu
Kevin J. Lawson VP Regulatory Affairs
60 Pineview Drive ● Buffalo, NY 14228-2120 USA 0 Toll free (800) 537-8378 ● tel. (716) 691-0091
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