ANA immunofluorescence versus profile-how well they perform in autoimmune diseases: an analysis of their clinical utility in a tertiary care centre

Petchiappan, Velammal; Guhan, Adityan; Selvam, Sumitra; Nagaprabu, V N

doi:10.18203/2320-6012.ijrms20183659

Cited by 7 publications

(10 citation statements)

References 5 publications

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“…In contrast, the specificity of the two tests was different, whereas ELISA was more specific than IFA. This observation was also previously reported using different ELISA and IFA kits [ 18 - 21 ]. These variables were reflected in the positive and negative predictive values of the two procedures, where IFA showed a higher negative predictive value but a lower positive predictive value for SLE compared to ELISA.…”

Section: Discussionsupporting

confidence: 88%

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Comparison of Indirect Immunofluorescence and Enzyme Immunoassay for the Detection of Antinuclear Antibodies

Khalifah¹,

Almansouri²,

Mowallad³

et al. 2022

Cureus

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Objective: The detection of autoantibodies directed toward nuclear antigens is one of the main criteria for the diagnosis of systemic lupus erythematosus (SLE), for which the most commonly used techniques are the enzyme immunoassay and immunofluorescence assay (IFA). However, the sensitivity and specificity of these tests vary between different techniques. Thus, in this study, we aimed to determine the superior method for detecting antinuclear antibodies (ANAs) and compare the accuracy of tests ordered by rheumatologists versus non-rheumatologists.Materials and methods: We compared the sensitivity and specificity of the two assays in 149 patients from a non-selected population, who were sent to the immunology laboratory of King Abdulaziz Medical City, Jeddah from 2018 to 2019.Results: The sensitivity and specificity of the indirect IFA were 77.78 % and 58.65%, respectively. The positive and negative predictive values of IFA for SLE were 44.87% and 85.92%, respectively. The sensitivity and specificity of the enzyme-linked immunosorbent assay (ELISA) were 77.78% and 80.77%, respectively. The negative and positive predictive values of ELISA for SLE were 63.64% and 89.36%, respectively. The highest number of false-positive IFA tests was requested by family physicians and the lowest was requested by rheumatologists. Conclusion: Our data show that IFA has a higher negative predictive value, while ELISA has a higher positive predictive value. The positive predictive value of the test can be improved by pre-selecting patients by specialist rheumatologists.

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Section: Discussionsupporting

confidence: 88%

“…In our study, the sensitivity of IFA and ELISA were similar (77.78%). Previous studies showed that the sensitivity of the two tests was close to each other [18][19][20][21]. In contrast, the specificity of the two tests was different, whereas ELISA was more specific than IFA.…”

Section: Discussionmentioning

confidence: 89%

Comparison of Indirect Immunofluorescence and Enzyme Immunoassay for the Detection of Antinuclear Antibodies

Khalifah¹,

Almansouri²,

Mowallad³

et al. 2022

Cureus

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“…All the study participants and ANA-IIF-positive patients had a 4% negative ANA profile EIA, which is significantly lower than the 27% found by Petchiappan et al [15]. This difference may be attributed to the fact that our study detected more antibodies that were missed by the ANA profile EIA kits used in Petchiappan et al Doing an ANA profile EIA directly may save time, but if the specific antigen is not included in the kit, it may not detect the antibodies.…”

Section: Discussioncontrasting

confidence: 66%

“…A study by Tarazi et al reported a negative ANA test in a diagnosed patient with cutaneous lupus Erythematosus [ 14 ]. Also, Petchiappan et al reported that 9 of 10 negative ANA-IIF SLE were positive for autoantibodies detected by ELISA [ 15 ].…”

Section: Introductionmentioning

confidence: 99%

Negative ANA-IIF in SLE patients: what is beyond?

et al. 2023

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The antinuclear antibody (ANA) test has high sensitivity in diagnosing and classifying systemic lupus erythematosus (SLE). Objectives To describe the immunological pattern of SLE patients through investigating specific antinuclear autoantibodies by enzyme dot immunoassay and studying their frequency in both positive and negative ANA indirect immunofluorescence assay (IIF) cases. Methods In a cross-sectional study, blood samples from 393 newly diagnosed SLE patients were analyzed using (IIF) on HEp-2 cells and ANA dot immunoassay by automated enzyme immunoassay (EIA) to detect 19 antibodies. Results Ninety-one percent of the patients are females; their mean age was 37 ± 12.28. Antinuclear antibody (ANA) was detected by IIF in 82.4% of cases, with 181 (46.1%) speckled and 167 (42.4%) homogeneous ANA patterns. The majority of patients (96%) demonstrated autoantibodies via EIA. Among the ANA-IIF-negative patients, 97.2% demonstrated autoantibodies. There was a significant difference in the frequency of certain autoantibodies between SLE patients with negative and positive ANA-IIF (1.44 0.73, 3.12 2.09, p = 0.00) respectively. Conclusion The results of analyzing 19 autoantibodies with the ANA staining pattern increased the significance of analyzing the immune profile even if IIF is negative when clinical symptoms strongly suggest SLE diagnosis. Certain autoantibodies may evade staining by the IFA approach while they are present in the patient’s serum, and they may not be detected by the ANA EIA profile if it does not contain that antigenic substrate. Key Points• Indirect immunofluorescence on Hep-2 is the conventional method for ANA detection and is regarded as the “gold standard” for testing in clinical practice for SLE.• In our study, ANA profile dot enzyme immunoassay (EIA)-based test was performed to evaluate 19 autoantibodies in SLE patients either positive or negative for ANA-IIF.• The presence of anti-dsDNA with ANA-IIF-negative serum in 32.4% of SLE patients provides evidence that not all anti-dsDNA antibodies are identified on standard HEp-2 substrates.• certain autoantibodies can evade staining by the ANA-IIF method despite being present in the SLE patient’s blood; this supports the ANA profile enzyme dot immunoassay as a more sensitive test.

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“…The study by Pachaiyappan V et al in South India also found that the most common pattern was nucleus speckled. 6 Where as, studies by Jacinth Angel etal. and Sebastian W et al in South India showed that nuclear homogenous pattern was the common pattern.…”

Section: Discussionmentioning

confidence: 98%

Antinuclear antibody patterns by indirect immunofluorescence test: An experience from a rural tertiary health centre of Puducherry, South India

Ranganathan

Rangasamy²,

Thiyagarajan³

et al. 2021

IJMR

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Anti-nuclear antibodies (ANA) are known to be associated with autoimmune diseases and most commonly with connective tissue diseases (CTD). Screening for ANA is the first investigation of choice in the diagnosis of autoimmune diseases and indirect immunofluorescence (IIF) test using Hep-2 cells has been considered the gold standard method. The prevalence of autoimmune diseases and hence the ANA positivity and ANA IIF patterns differ in various geographical regions. This study was aimed to estimate the ANA positivity rates among patients with suspected or confirmed auto immune diseases and to determine the common ANA IIF patterns in the positive samples.: This is a retrospective study including 235 patients with either clinical suspicion of, or suffering from autoimmune diseases attending a tertiary health care hospital in Puducherry, South India, between November 2018 and April 2020. ANA IIF test (Hep 20-10/ primate liver, Euroimmun, Germany) was performed with 1:100 dilutions of patient serum.The ANA positivity rate was 14.5% (34 out of 235 samples were positive). ANA was positive in 10 (13.3%) males and 24 (15%) females. The mean age of ANA positive patients were 43.4 years.The most common pattern was nucleus fine granular (n=8) followed by cytoplasm fine granular (n=7) and centromere (n=4).A significant ANA positivity rate was found in our study and a detailed clinical study has to be conducted to draw further conclusions.

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ANA immunofluorescence versus profile-how well they perform in autoimmune diseases: an analysis of their clinical utility in a tertiary care centre

Cited by 7 publications

References 5 publications

Comparison of Indirect Immunofluorescence and Enzyme Immunoassay for the Detection of Antinuclear Antibodies

Comparison of Indirect Immunofluorescence and Enzyme Immunoassay for the Detection of Antinuclear Antibodies

Negative ANA-IIF in SLE patients: what is beyond?

Antinuclear antibody patterns by indirect immunofluorescence test: An experience from a rural tertiary health centre of Puducherry, South India

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