(266 days)
ImmuGlo ANTINUCLEAR ANTIBODY (ANA) TEST (HEP-2 CELLS)
Not Found
No
The summary describes a traditional indirect immunofluorescence assay kit and its performance characteristics. There is no mention of automated image analysis, pattern recognition, or any computational methods that would typically involve AI/ML. The device relies on visual interpretation of fluorescence patterns by trained personnel.
No.
The device is used to detect anti-nuclear antibodies (ANA) as an aid in diagnosing systemic rheumatic diseases, not for treatment.
Yes
Explanation: The "Intended Use / Indications for Use" section explicitly states, "The ImmuGlo HEp-2 Elite IFA is intended for use as an aid in the diagnosis of systemic rheumatic diseases in conjunction with other clinical and laboratory findings." This clearly indicates its role in the diagnostic process.
No
The device is a laboratory test kit that utilizes physical reagents (HEp-2 cells, serum) and requires fluorescence microscopy for analysis. It is not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The "Intended Use / Indications for Use" section explicitly states that the device is an "indirect immunofluorescence antibody test for the qualitative or semi-quantitative detection of anti-nuclear antibodies (ANA) of the IgG isotype in human serum". This indicates that the device is used to test a sample taken from the human body (serum) in vitro (outside the body) to provide information about a medical condition (presence of ANA).
- Device Description: The description further clarifies that it utilizes "human serum" and "HEp-2 cells as a substrate", reinforcing the in vitro nature of the test.
- Performance Studies: The document details performance studies conducted on "human serum" samples, including method comparison, pattern agreement, precision, interference, and a clinical study using "clinical samples". These are all typical evaluations for an IVD.
- Aid in Diagnosis: The intended use states it is "intended for use as an aid in the diagnosis of systemic rheumatic diseases". This is a common purpose for IVD devices.
Therefore, based on the provided information, the ImmuGlo HEp-2 Elite IFA clearly fits the definition of an In Vitro Diagnostic device.
N/A
Intended Use / Indications for Use
The ImmuGlo HEp-2 Elite IFA is an indirect immunofluorescence antibody test for the qualitative or semi-quantitative detection of anti-nuclear antibodies (ANA) of the IgG isotype in human serum utilizing standard HEp-2 cells and engineered HEp-2 cells as a substrate. The ImmuGlo HEp-2 Elite IFA is intended for use as an aid in the diagnosis of systemic rheumatic diseases in conjunction with other clinical and laboratory findings.
Product codes
DHN
Device Description
Antinucles (ANA) target various nuclear and cellular components and are highly prevalent in systemic autoimmune connective tissue disorders. ANA can be detect immunofluorescence assay (IFA) using HEp-2 cell substrates. Such assys aid in the diagnosis of systemic lupus erythematosus (SLE) and other including mixed connective tissue disease, Sjögren's syndrome, systemic sclerosis, polymyositis, drug induced lupus, as well as cases of autoimmune hepatitis. ANA occur in about 95% of SLE patients. HEp-2 cell lines present a variety of autoantigens that are not found in solid phase assays of ANA. Solid phase assays developed on various platforms differ in their and specificity raising uncertainties regarding the interpretation of results. Due to the limitations of solid phase assays, the American College of Rheumatology (ACR) considers IFA using a HEp-2 cell substrate as the gold standard for ANA testing.
The HEp-2-lFA method is able to report both ANA pattern and titer for the test specimen. Various groups have worked to improve awareness of ANA patterns detected by HEp-2, criteria for accurate interpretation of ANA patterns. Much of this work has been compiled in the International Consensus on Antibody Patterns (CAP) report recommending a standardized nomenclature and reporting format for IFA patterns.
The major nuclear patterns observed on HEp-2 substrates include Homogeneous (AC-1), Speckled (AC-2, AC-4, AC-5), Centromere (AC-3), Discrete Nuclear Dots (AC-6, AC-7) and Nucleolar (AC-8, AC-9 and AC-10) patterns. A critical development in recent years is the definition of a frequently reported pattern termed "Dense Fine Speckled" that has been assigned the anti-cell pattern number AC-2 and recommended as a competent level reporting parameter by CAP. The major autoantigenic target associated with the DFS/AC-2 pattern, also referred to as the DF570 pattern, is a 70 kDa antigen known as LEDGF (Lens Epithelium Derived Growth Factor) or PSP1 gene product. These autoantibodies were initially reported to occur in sera from patients with positive ANA tests but no clinical evidence of systemic alsease (SARD) and do not have a distinct clinical association. The prevalence of DF570 autoantibodies, methods used to screen/confirm the DFS/AC-2 pattern (i.e. IFA or solidphase) and implications of inaccurate interpretation of DF570/DFS/AC-2 pattern have been presented in literature. The DFS/DFS70/AC-2 pattern may be difficult to read and can be mis-interpreted as homogeneous (associated with DNA, histones and nucleosomes), fine speckled, or a mix of homogeneous and fine speckled patterns, all of which are associated with systemic autoimmune diseases of the DFS70 pattern and the lack of commercially available assays that aid in the DFS/DF570/AC-2 pattern from mixed disease associated patterns increases the need for a method that may aid in distinction between ANA patterns at the screening stage without compromising the inherent advantages of the HEp-2 IFA method. The ICAP committee recommends all clinical labs report the DFS70 pattern.
HEp-2 Elite sides provided with this kit contain a 1:9 mixture of standard HEp-2 cells devoid of functional copies of PSP2 gene (UniGene Gene ID: 1116). The engineered HEp-2 cels are able to detect all ANA specificities with the exception of the DF570/AC-2 pattern resulting from autoantibodies associated with PSP2. In DF570 positive reactions standard HEp-2 cells provide a positive reaction while engineered HEp-2 cells do not. The HEp-2 Elite substrate thereby provides additionality to aid in discriminating homogeneous, speckled, and dense fine speckled (DFS70/AC-2) patterns during the screening phase.
HEp-2 Elite IFA is an indirect immunofluorescence antibody test for the qualitative detection of anti-nuclear antibodies (ANA) of the IgG isotype in human serum utilizing standard HEp-2 cells as a substrate. The ImmuGlo™ HEp-2 Elite FA is intended for use as an aid in the diseases in conjunction with other clinical and laboratory findings.
Features: Both kits use HEp-2 cells in and positive controls, serum diluent, wash solution, lgG conjugate and mounting medium to detect ANA IgG antibodies. Both kits use 1:40 screening dilutions and 30 minute incubation steps and slides are read on a fluorescence microscope. The ImmuGlo HEp-2 cells mixed with engineered HEp-2 with the PSP1 gene knocked out in 1:9 ratio while the ImmuGlo ANTINUCLEAR ANTIBODY (ANA) TEST utilizes only standard HEp-2 cells.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
fluorescence microscope
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
Not Found
Description of the test set, sample size, data source, and annotation protocol
Non-clinical Tests:
Method Comparison: Both kits were tested with well-characterized ANA associated disease controls.
No specific details on data source or annotation protocol. Sample size is 591 total (298 positive, 293 negative).
Pattern Agreement Study:
An additional study was designed to detern agreement using 243 samples with known ANA patterns. These samples were randomized and sides provided to 3 independent evaluators. The readers were blinded to each others' results, as well as ever the samples. The data was analyzed by comparing Elite vs Predicate by read (at least 2 of 3 readers). In the case of mixed patterns where more than one pattern was observed, the dominant or primary pattern was analyzed.
No specific details on data source or annotation protocol.
Clinical Study:
Sets of clinical samples were tested on the Included 256 AM associated disease samples and 497 autoimmune and infectious disease controls.
ANA associated diseases used for calculation of sensitis, Antiphospholipid syndrome, Mixed connective tissue disorder, Myositis, Siögren's syndrome, Systemic sclerosis and Drug induced lupus erythematosus.
Disease Control samples (non-ANA associated diseases) used for calculation of specificity included Celiac disease, Pernicious anemia, Primary biliary cirrhosis, Rheumatoid arthritis, Autoimmune thyroid disease, Ulcerative colitis, Raynaud's syndrome, Psoriasis, Atopic dermatitis, Colorectal cancer, Ovarian cancer, Breast cancer, Prostate cancer, Cytomegalovirus, Hepatitis C, Herpes simplex virus, Lyme disease, Mononucleosis, Rubella, Syphilis, Toxoplasmosis, Human immunodeficiency virus and Hepattis B.
Normal human sera and DF570 suspected samples were excluded in sensitivity/specificity calculations.
No specific details on data source or annotation protocol.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Non-clinical Tests:
Method Comparison:
Type: Comparison study against predicate device.
Sample size: 591 (298 positive, 293 negative).
Key results: Positive % Agreement 99.7% (95% CI 97.8% - 100.0%), Negative % Agreement 98.3% (95% CI 95.8% - 99.4%), Overall % Agreement 99.3% (95% CI 97.8% - 99.5%).
Pattern Agreement Study:
Type: Reader agreement study.
Sample size: 243 samples.
Key results (Consensus (2/3 Readers)):
All: PPA 88.9% (83.1-92.9%), NPA 76.2% (63.5-85.6%).
DFS(70): PPA 100.0% (84.0-100.0%), NPA 93.1% (88.6-95.9%).
Homogeneous: PPA 76.4% (62.7-86.3%), NPA 100.0% (97.5-100.0%).
Speckled: PPA 97.5% (85.3-99.9%), NPA 97.0% (93.4-98.8%).
Precision:
Repeatability:
Type: Repeatability study.
Sample size: 8 negative and 17 positive samples, tested in 6 replicates in 10 runs per day for 5 days, resulting in 60 replicates for each sample read by 2 operators for a total n of 120.
Key results: All samples met the 90% acceptance criteria for qualitative agreement (positive), fluorescence intensity agreement (± 1 fluorescence value within one run) and pattern agreement.
Reproducibility:
Type: Reproducibility study.
Sample size: 10 positive samples tested in triplicate in 10 runs (2 runs per day for 5 days) at three different sites, resulting in 90 replicates for each sample.
Key results: Overall agreement was 99.75%. The acceptance criteria for these studies was 95% or greater and rs within ± 1 fluorescence value, ± 1 titer with consent.
Specific sample results for pattern, titer, and fluorescence intensity agreement are provided in a table.
Interference:
Type: Interference study.
Key results: No significant interference was demonstrated for hemoglobin, unconjugated bilirubin, rheumatoid factor, triglycerides, naproxen, ibuprofen, prednisone, mycophernyate mofetil, hydroxychloroquine, cyclophosphamide, rituximab, and Belimumab at specified levels.
Clinical Study:
Type: Clinical sample testing.
Sample size: 256 ANA associated disease samples and 497 autoimmune and infectious disease controls.
Key results: ANA Associated Disease Sensitivity: 78.1% (95% C.I. 72.5-82.9%), ANA Associated Disease Specificity: 75.1% (95% C.I. 71.0-78.7%).
Stability:
Type: Accelerated and real-time stability testing.
Key results: Accelerated stability testing on 3 lots showed no drop in fluorescence over 30 days, supporting predicate shelf life of 18 months. Real-time and open kit stability is currently 18 months for 2 lots and 9 for a third, which all continue to meet AQL.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Positive % Agreement 99.7% (95% CI 97.8% - 100.0%)
Negative % Agreement 98.3% (95% CI 95.8% - 99.4%)
Overall % Agreement 99.3% (95% CI 97.8% - 99.5%)
ANA Associated Disease Sensitivity: 78.1% (95% C.I. 72.5-82.9%)
ANA Associated Disease Specificity: 75.1% (95% C.I. 71.0-78.7%)
Pattern Agreement Study (Consensus (2/3 Readers)):
All: PPA 88.9% (83.1-92.9%)
All: NPA 76.2% (63.5-85.6%)
DFS(70): PPA 100.0% (84.0-100.0%)
DFS(70): NPA 93.1% (88.6-95.9%)
Homogeneous: PPA 76.4% (62.7-86.3%)
Homogeneous: NPA 100.0% (97.5-100.0%)
Speckled: PPA 97.5% (85.3-99.9%)
Speckled: NPA 97.0% (93.4-98.8%)
Predicate Device(s)
ImmuGlo ANTINUCLEAR ANTIBODY (ANA) TEST (HEP-2 CELLS)
Reference Device(s)
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 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).
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Image /page/0/Picture/0 description: The image contains the logos of the Department of Health & Human Services and the Food and Drug Administration (FDA). The Department of Health & Human Services logo is on the left, and the FDA logo is on the right. The FDA logo includes the words "FDA U.S. FOOD & DRUG ADMINISTRATION" in blue text.
June 05, 2018
IMMCO Diagnostics, Inc. Kevin Lawson Chief Regulatory Officer 60 Pineview Drive Buffalo, New York 14228
Re: K172745
Trade/Device Name: ImmuGlo HEp-2 Elite IFA Regulation Number: 21 CFR 866.5100 Regulation Name: Antinuclear antibody immunological test system Regulatory Class: Class II Product Code: DHN Dated: September 8, 2017 Received: September 12, 2017
Dear Kevin 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 Part 801 and Part 809); medical device 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
1
Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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 comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely.
Leonthena R. Carrington -S
Lea Carrington Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
2
Indications for Use
510(k) Number (if known) K172745
Device Name ImmuGlo HEp-2 Elite IFA
Indications for Use (Describe)
The ImmuGlo HEp-2 Elite IFA is an indirect immunofluorescence antibody test for the qualitative or semi-quantitative detection of anti-nuclear antibodies (ANA) of the IgG isotype in human serum utilizing standard HEp-2 cells and engineered HEp-2 cells as a substrate. The ImmuGlo HEp-2 Elite IFA is intended for use as an aid in the diagnosis of systemic rheumatic diseases in conjunction with other clinical and laboratory 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 circles on the left side, followed by the company name in blue, with the word "DIAGNOSTICS" in green below it. Underneath the company name, it says "A Trinity Biotech Company" in a smaller font.
ImmuGlo HEp-2 Elite IFA 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: | 6-5-2018 |
| Device Name: | ImmuGlo™ HEp-2 Elite IFA |
| Common Name: | HEp-2 IFA |
| Product Code: | Antinuclear Antibody, Indirect Immunofluorescent, Antigen, Control [DHN |
| Substantially Equivalent to: | ImmuGlo ANTINUCLEAR ANTIBODY (ANA) TEST (HEP-2 CELLS) |
General Description: Antinucles (ANA) target various nuclear and cellular components and are highly prevalent in systemic autoimmune connective tissue disorders. ANA can be detect immunofluorescence assay (IFA) using HEp-2 cell substrates. Such assys aid in the diagnosis of systemic lupus erythematosus (SLE) and other including mixed connective tissue disease, Sjögren's syndrome, systemic sclerosis, polymyositis, drug induced lupus, as well as cases of autoimmune hepatitis. ANA occur in about 95% of SLE patients. HEp-2 cell lines present a variety of autoantigens that are not found in solid phase assays of ANA. Solid phase assays developed on various platforms differ in their and specificity raising uncertainties regarding the interpretation of results. Due to the limitations of solid phase assays, the American College of Rheumatology (ACR) considers IFA using a HEp-2 cell substrate as the gold standard for ANA testing.
The HEp-2-lFA method is able to report both ANA pattern and titer for the test specimen. Various groups have worked to improve awareness of ANA patterns detected by HEp-2, criteria for accurate interpretation of ANA patterns. Much of this work has been compiled in the International Consensus on Antibody Patterns (CAP) report recommending a standardized nomenclature and reporting format for IFA patterns.
The major nuclear patterns observed on HEp-2 substrates include Homogeneous (AC-1), Speckled (AC-2, AC-4, AC-5), Centromere (AC-3), Discrete Nuclear Dots (AC-6, AC-7) and Nucleolar (AC-8, AC-9 and AC-10) patterns. A critical development in recent years is the definition of a frequently reported pattern termed "Dense Fine Speckled" that has been assigned the anti-cell pattern number AC-2 and recommended as a competent level reporting parameter by CAP. The major autoantigenic target associated with the DFS/AC-2 pattern, also referred to as the DF570 pattern, is a 70 kDa antigen known as LEDGF (Lens Epithelium Derived Growth Factor) or PSP1 gene product. These autoantibodies were initially reported to occur in sera from patients with positive ANA tests but no clinical evidence of systemic alsease (SARD) and do not have a distinct clinical association. The prevalence of DF570 autoantibodies, methods used to screen/confirm the DFS/AC-2 pattern (i.e. IFA or solidphase) and implications of inaccurate interpretation of DF570/DFS/AC-2 pattern have been presented in literature. The DFS/DFS70/AC-2 pattern may be difficult to read and can be mis-interpreted as homogeneous (associated with DNA, histones and nucleosomes), fine speckled, or a mix of homogeneous and fine speckled patterns, all of which are associated with systemic autoimmune diseases of the DFS70 pattern and the lack of commercially available assays that aid in the DFS/DF570/AC-2 pattern from mixed disease associated patterns increases the need for a method that may aid in distinction between ANA patterns at the screening stage without compromising the inherent advantages of the HEp-2 IFA method. The ICAP committee recommends all clinical labs report the DFS70 pattern.
HEp-2 Elite sides provided with this kit contain a 1:9 mixture of standard HEp-2 cells devoid of functional copies of PSP2 gene (UniGene Gene ID: 1116). The engineered HEp-2 cels are able to detect all ANA specificities with the exception of the DF570/AC-2 pattern resulting from autoantibodies associated with PSP2. In DF570 positive reactions standard HEp-2 cells provide a positive reaction while engineered HEp-2 cells do not. The HEp-2 Elite substrate thereby provides additionality to aid in discriminating homogeneous, speckled, and dense fine speckled (DFS70/AC-2) patterns during the screening phase.
Intended Use: The ImmuGlo™ HEp-2 Elite IFA is an indirect immunofluorescence antibody test for the qualitative detection of anti-nuclear antibodies (ANA) of the IgG isotype in human serum utilizing standard HEp-2 cells as a substrate. The ImmuGlo™ HEp-2 Elite FA is intended for use as an aid in the diseases in conjunction with other clinical and laboratory findings.
Similarities and Differences: Both kits use HEp-2 cells in and positive controls, serum diluent, wash solution, lgG conjugate and mounting medium to detect ANA IgG antibodies. Both kits use 1:40 screening dilutions and 30 minute incubation steps and slides are read on a fluorescence microscope. The ImmuGlo HEp-2 cells mixed with engineered HEp-2 with the PSP1 gene knocked out in 1:9 ratio while the ImmuGlo ANTINUCLEAR ANTIBODY (ANA) TEST utilizes only standard HEp-2 cells.
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/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 dots on the left, followed by the company name "immco" in blue, with "DIAGNOSTICS" in green underneath. Below the company name, it says "A Trinity Biotech Company" in a smaller font.
Non-clinical Tests:
Method Comparison: Both kits were tested with well-characterized ANA associated disease controls.
| Predicate | ||||
|---|---|---|---|---|
| Positive | Negative | Total | ||
| Hep-2 | Positive | 297 | 5 | 302 |
| Elite | Negative | 1 | 288 | 289 |
| IFA | Total | 298 | 293 | 591 |
| Positive % Agreement 99.7% (95% CI 97.8% - 100.0%) | ||||
| Negative % Agreement 98.3% (95% CI 95.8% - 99.4%) | ||||
| Overall % Agreement 99.3% (95% CI 97.8% - 99.5%) |
Qualitative Agreement (Pos/ Neg)
Pattern Agreement Study
An additional study was designed to detern agreement using 243 samples with known ANA patterns. These samples were randomized and sides provided to 3 independent evaluators. The readers were blinded to each others' results, as well as ever the samples. The data was analyzed by comparing Elite vs Predicate by read (at least 2 of 3 readers). In the case of mixed patterns where more than one pattern was observed, the dominant or primary pattern was analyzed.
| Elite vs Predicate | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Pattern | Reader 1 | Reader 2 | Reader 3 | Consensus | |||||
| (2/3 Readers) | |||||||||
| n | % (95% CI) | n | % (95% CI) | n | % (95% CI) | n | % (95% CI) | ||
| All | PPA | 162 | 98.8% | ||||||
| (95.1-99.8%) | 144 | 95.8% | |||||||
| (90.8-98.3%) | 197 | 95.4% | |||||||
| (91.2-97.8%) | 180 | 88.9% | |||||||
| (83.1-92.9%) | |||||||||
| NPA | 81 | 81.5% | |||||||
| (71.0-88.9%) | 99 | 67.7% | |||||||
| (57.4-76.5%) | 46 | 78.2% | |||||||
| (63.2-88.5%) | 63 | 76.2% | |||||||
| (63.5-85.6%) | |||||||||
| DFS(70) | PPA | 26 | 100.0% | ||||||
| (84.0-100.0%) | 21 | 95.2% | |||||||
| (74.1-99.7%) | 42 | 100.0% | |||||||
| (89.6-100.0%) | 26 | 100.0% | |||||||
| (84.0-100.0%) | |||||||||
| NPA | 217 | 94.4% | |||||||
| (89.7-96.6%) | 222 | 89.6% | |||||||
| (84.7-93.2%) | 201 | 99.5% | |||||||
| (96.8-100.0%) | 217 | 93.1% | |||||||
| (88.6-95.9%) | |||||||||
| Homogeneous | PPA | 54 | 81.5% | ||||||
| (68.1-90.3%) | 57 | 68.4% | |||||||
| (54.6-79.7%) | 49 | 91.8% | |||||||
| (79.5-97.4%) | 55 | 76.4% | |||||||
| (62.7-86.3%) | |||||||||
| NPA | 189 | 100.0% | |||||||
| (97.5-100.0%) | 186 | 98.9% | |||||||
| (95.7-99.8%) | 194 | 98.5% | |||||||
| (95.2-99.6%) | 188 | 100.0% | |||||||
| (97.5-100.0%) | |||||||||
| Speckled | PPA | 45 | 88.9% | ||||||
| (75.2-95.8%) | 47 | 72.3% | |||||||
| (57.1-83.9%) | 53 | 83.0% | |||||||
| (69.7-91.5%) | 40 | 97.5% | |||||||
| (85.3-99.9%) | |||||||||
| NPA | 198 | 99.9% | |||||||
| (96.0-99.8%) | 196 | 98.0% | |||||||
| (94.5-99.3%) | 190 | 96.3% | |||||||
| (92.3-98.4%) | 203 | 97.0% | |||||||
| (93.4-98.8%) |
NOTE: Above data depicts primary pattern only. Results will differ if secondary pattern is included. n values depict samples positive or negative by the predicate device.
OBSERVED PREVALENCE
Tests for nuclear antibodies are used to screen other immunologic disturbances. Sets of clinical samples were tested on the HEp-2 Elite IFA. Results demonstrating prevalence in the populations for this study are provided below.
| Condition | n Tested | n Positive | % Positive |
|---|---|---|---|
| ANA-associated autoimmune disease | 256 | 200 | 78.1% |
| Autoimmune hepatitis | 13 | 6 | 46.2% |
| Antiphospholipid syndrome | 15 | 7 | 46.7% |
| Mixed connective tissue disease | 10 | 7 | 70.0% |
| Myositis | 36 | 16 | 44.4% |
| Sjögren's | 65 | 62 | 95.4% |
60 Pineview Drive Toll free (800) 537-8378 fax (716) 691-0466
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Image /page/5/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 a smaller font, and below that is the text "A Trinity Biotech Company".
| Systemic lupus erythematosus | 78 | 74 | 94.9% |
|---|---|---|---|
| Systemic sclerosis | 27 | 17 | 63.0% |
| Drug-induced lupus | 12 | 11 | 91.7% |
| Non-ANA-associated autoimmune disease | 254 | 90 | 35.4% |
| Celiac | 10 | 1 | 10.0% |
| Crohn's | 25 | 6 | 24.0% |
| Pernicious anemia | 20 | 1 | 5.0% |
| Primary biliary cirrhosis | 25 | 17 | 68.0% |
| Rheumatoid arthritis | 87 | 52 | 68.0% |
| Autoimmune thyroiditis | 20 | 2 | 10.0% |
| Ulcerative colitis | 23 | 6 | 26.1% |
| ANCA Vasculitis | 20 | 1 | 5.0% |
| Raynaud's syndrome | 4 | 2 | 50.0% |
| Psoriasis | 20 | 2 | 10.0% |
| Inflammatory disease | 13 | 2 | 15.4% |
| Atopic dermatitis | 13 | 2 | 15.4% |
| Cancer | 57 | 3 | 5.3% |
| Colorectal cancer | 10 | 1 | 10.0% |
| Ovarian cancer | 10 | 0 | 0.0% |
| Pancreatic cancer | 9 | 0 | 0.0% |
| Breast cancer | 8 | 1 | 12.5% |
| Prostate cancer | 20 | 1 | 5.0% |
| Infectious Disease | 173 | 29 | 16.8% |
| Cytomegalovirus | 20 | 1 | 5.0% |
| Hepatitis C | 11 | 4 | 36.4% |
| Herpes simplex virus | 20 | 2 | 10.0% |
| Lyme | 12 | 6 | 50.0% |
| Mononucleosis | 10 | 2 | 20.0% |
| Rubella | 20 | 4 | 20.0% |
| Syphilis | 20 | 2 | 10.0% |
| Toxoplasmosis | 20 | 5 | 25.0% |
| HIV | 20 | 2 | 10.0% |
| HBV | 20 | 1 | 5.0% |
| Normal human serum | 128 | 7 | 5.5% |
Cross Reactivity See Observed Prevalence section above.
Precision
Repeatability
To assess repeatability of the ImmuGlo™ HEp-2 Elite IFA and a manual microscope 8 negative and 17 positive samples with various patterns and intensities were tested in 6 replicates in 10 runs per day for 5 days), resulting in 60 replicates for each sample read by 2 operators for a total n of 120. All samples met the 90% acceptance criteria for qualitative agreement (positive), fluorescence intensity agreement (± 1 fluorescence value within one run) and pattern agreement.
Reproducibility
In another study, the ImmuGl" HEp-2 Elte IFA and a manual microscope 10 positive sample with various patterns and intensities tested in triplicate in 10 runs (2 runs per day for 5 days) at three different sites, resulting in 90 replicates for each sample. % pattern agreement, % titer agreement and % fluorescenent results are provided below. The AQL for these studies was ± 1 fluorescence value, ± 1 titer and a consensus pattern agreement. The acceptance criteria for these studies was 95% or greater and rs within ± 1 fluorescence value, ± 1 titer with consent. The two samples that did not meet AQL were the result of subjective reader error. Overall agreement was 99.75%. Samples 2 and 9 demonstrated secondary pattern and intensity for these samples. No report of a secondary pattern is considered agreement for all other samples.
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| Sample | Pattern | Endpoint
Titer | Fluorescence
Intensity | Pattern
Agreement | Titer
Agreement | Fluorescence
Intensity
Agreement |
|--------|--------------|-------------------|---------------------------|----------------------|--------------------|----------------------------------------|
| 1 | DFS70 | 80 | 2+ | 100% | 100% | 100% |
| 2* | Cytoplasmic | 80 | 2+ | 100% | 100% | 100% |
| | Nuclear dots | 80 | 2+ | 100% | 100% | 100% |
| 3 | Speckled | 80 | 2+ | 100% | 100% | 100% |
| 4 | Centromere | 80 | 2+ | 100% | 100% | 100% |
| 5 | Speckled | 80 | 2+ | 100% | 100% | 100% |
| 6 | Homogeneous | 40 | 1+ | 93% | 100% | 100% |
| 7 | DFS70 | 80 | 2+ | 100% | 100% | 100% |
| 8 | Homogeneous | 160 | 3+ | 100% | 93% | 100% |
| 9* | Cytoplasmic | 1280 | 3+ | 100% | 100% | 100% |
| | Nuclear dots | 1280 | 4+ | 100% | 100% | 100% |
| 10 | Nucleolar | 1280 | 4+ | 100% | 100% | 100% |
| 11 | Negative | 0 | 0 | 100% | 100% | 100% |
- Samples 2 and 9 are mixed pattern specificity, primary pattern is listed first.
Interference
Interference was studied by mixing sera with potentially interfering serum samples and studying deviation from expected results. No significant interference was demonstrated for the following substances at the levels indicated: hemoglobin (2 g/L), unconjugated bilirubin (342 µmo/(), rheumatoid factor (13 mmo/4), triglycerides (3.7 mmo/4), naproxen (2170 µmo/4), ibuprofen (2425 umol/L), prednisone (0.84 µmo/L), mycophernyate mofetil (14.01 µg/ml), hydroxychloroquine (388 ng/ml), cyclophosphamide (1437 µmol/L), rituximab (1.143 mg/ml) and Belimumab (0.939 mg/ml).
Clinical Study: Sets of clinical samples were tested on the Included 256 AM associated disease samples and 497 autoimmune and infectious disease controls.
ANA Associated Disease Sensitivity: 78.1% (95% C.I. 72.5-82.9%) ANA Associated Disease Specificity: 75.1% (95% C.I. 71.0-78.7%).
ANA associated diseases used for calculation of sensitis, Antiphospholipid syndrome, Mixed connective tissue disorder, Myositis, Siögren's syndrome, Systemic sclerosis and Drug induced lupus erythematosus. Disease Control samples (non-ANA associated diseases) used for calculation of specificity included Celiac disease, Pernicious anemia, Primary biliary cirrhosis, Rheumatoid arthritis, Autoimmune thyroid disease, Ulcerative colitis, Raynaud's syndrome, Psoriasis, Atopic dermatitis, Colorectal cancer, Ovarian cancer, Breast cancer, Prostate cancer, Cytomegalovirus, Hepatitis C, Herpes simplex virus, Lyme disease, Mononucleosis, Rubella, Syphilis, Toxoplasmosis, Human immunodeficiency virus and Hepattis B. Normal human sera and DF570 suspected samples were excluded in sensitivity/specificity calculations.
Stability: All reagents for this kit are the sredicate device except for the slide, so accelerated stability testing was performed on the slide to ensure no change in shelf life. Accelerated stability testing on 3 lots stored at 37℃ for up to 30 days showed no drop in fluorescence over the course of the experiment supporting the predicate shelf life of 18 months. Real time and open kit stability is currently 18 months for 2 lots and 9 for a third, which all continue to meet AQL.
Ken Burns
Kevin J. Lawson Chief Regulatory Officer
60 Pineview Drive Toll free (800) 537-8378 · tel. (716) 691-0091 · · fax (716) 691-0466
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