A Multicenter Analysis of Subjectivity of Indirect Immunofluorescence Test in Antinuclear Antibody Screening

Faraşat, Vildan Turan; Ecemiş, Talat; Doğan, Yavuz; Şener, Aslı Gamze; Ece, Gülfem; Dündar, Pınar Erbay; Şanlıdağ, Tamer

doi:10.5606/archrheumatol.2019.7310

Cited by 6 publications

(4 citation statements)

References 17 publications

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“…However, in this study, positive anti-SSA or anti-Mi2 results may not always be accompanied by a positive speckled pattern. We believe that the methods of ANA detection and ANA-specific antibody detection differ, and the major challenge and limitation of ANA-IIF testing is that it requires competent and experienced interpreters and is subjective ( 27 ), which may explain the above results.…”

Section: Discussionmentioning

confidence: 99%

A single-centre study on abnormal antinuclear antibodies in children caused by intravenous infusion of gamma globulin

Xu,

Zhou,

Zhang

et al. 2024

Front. Immunol.

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ObjectiveTo clarify the impact of intravenous infusion of gamma globulin (IVIg) on antinuclear antibodies (ANAs) in children.MethodsA retrospective analysis was performed on the data of children with nonspecific autoantibody-related diseases whose antinuclear antibody (ANA) and autoantibody profiles were detected in our hospital from January to March 2022. A total of 108 patients with a clear history of IVIg infusion within 28 days composed the IVIg group, and 1201 patients without a history of IVIg infusion composed the non-IVIg group.ResultsAll patients in the IVIg group had either positive ANAs or positive autoantibodies. Anti-SSA, anti-Ro52 and anti-AMA Mi2 were the top three autoantibodies in the IVIg group. The proportions of patients who were positive for either of these three autoantibodies in the IVIg group were significantly greater than those in the non-IVIg group (all P<0.5). Spearman correlation analysis revealed that the signal intensities of anti-SSA and anti-Ro52 were negatively correlated with the number of days of ANA detection after IVIg infusion (P<0.05). Multiple logistic analyses revealed that a greater total dosage of IVIg, greater IVIg per kilogram of body weight, and fewer ANA detection days after IVIg infusion were independent risk factors for positive anti-SSA and anti-Ro52 results.ConclusionsIt is recommended that if rheumatic diseases are suspected, ANA detection should be carried out beforeIVIg infusion. But for patients who are positive for at least one of these three autoantibodies after IVIg infusion, doctors should first consider adoptive antibodies.

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Section: Discussionmentioning

confidence: 99%

A single-centre study on abnormal antinuclear antibodies in children caused by intravenous infusion of gamma globulin

Xu,

Zhou,

Zhang

et al. 2024

Front. Immunol.

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“…A recent multicenter analysis to evaluate the interpretation of HEp-2 IFA reported significant differences among laboratories in terms of qualitative results, patterns, and titers, particularly at low levels and in those with speckled patterns [ 24 ]. HEp-2 IFA titer determinations have been reported to have clinical significance in predicting risk for disease (healthy vs. disease) as well as association with specific autoantibodies [ 25 – 29 ].…”

Section: Discussionmentioning

confidence: 99%

The antinuclear antibody HEp-2 indirect immunofluorescence assay: a survey of laboratory performance, pattern recognition and interpretation

Tebo

Schmidt

Kadkhoda

et al. 2021

Autoimmun Highlights

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Background To evaluate the interpretation and reporting of antinuclear antibodies (ANA) by indirect immunofluorescence assay (IFA) using HEp-2 substrates based on common practice and guidance by the International Consensus on ANA patterns (ICAP). Method Participants included two groups [16 clinical laboratories (CL) and 8 in vitro diagnostic manufacturers (IVD)] recruited via an email sent to the Association of Medical Laboratory Immunologists (AMLI) membership. Twelve (n = 12) pre-qualified specimens were distributed to participants for testing, interpretation and reporting HEp-2 IFA. Results obtained were analyzed for accuracy with the intended and consensus response for three main categorical patterns (nuclear, cytoplasmic and mitotic), common patterns and ICAP report nomenclatures. The distributions of antibody titers of specimens were also compared. Results Laboratories differed in the categorical patterns reported; 8 reporting all patterns, 3 reporting only nuclear patterns and 5 reporting nuclear patterns with various combinations of other patterns. For all participants, accuracy with the intended response for the categorical nuclear pattern was excellent at 99% [95% confidence interval (CI): 97–100%] compared to 78% [95% CI 67–88%] for the cytoplasmic, and 93% [95% CI 86%–100%] for mitotic patterns. The accuracy was 13% greater for the common nomenclature [87%, 95% CI 82–90%] compared to the ICAP nomenclature [74%, 95% CI 68–79%] for all participants. Participants reporting all three main categories demonstrated better performances compared to those reporting 2 or less categorical patterns. The average accuracies varied between participant groups, however, with the lowest and most variable performances for cytoplasmic pattern specimens. The reported titers for all specimens varied, with the least variability for nuclear patterns and most titer variability associated with cytoplasmic patterns. Conclusions Our study demonstrated significant accuracy for all participants in identifying the categorical nuclear staining as well as traditional pattern assignments for nuclear patterns. However, there was less consistency in reporting cytoplasmic and mitotic patterns, with implications for assigning competencies and training for clinical laboratory personnel.

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“…A recent multicenter analysis to evaluate the interpretation of HEp-2 IFA reported significant differences among laboratories in terms of qualitative results, patterns, and titers, particularly at low levels and in those with speckled patterns [24]. HEp-2 IFA titer determinations have been reported to have clinical significance in predicting risk for disease (healthy vs. disease) as well as association with specific autoantibodies [25][26][27][28][29].…”

Section: Discussionmentioning

confidence: 99%

Reporting of Antinuclear Antibody HEp-2 Indirect Immunofluorescence Assay: A Performance Survey

Tebo¹,

Schmidt²,

Kadkhoka³

et al. 2020

Preprint

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Background: To evaluate the interpretation and reporting of antinuclear antibodies (ANA) by indirect immunofluorescence assay (IFA) using HEp-2 substrate based on common practice and guidance by the International Consensus on ANA patterns (ICAP).Method: Participants included two groups [16 clinical laboratories (CL) and 8 in vitro diagnostic manufacturers (IVD)] recruited via an email sent to the Association of Medical Laboratory Immunologists (AMLI) membership. Twelve (n=12) pre-qualified specimens were distributed to participants for testing, interpretation and reporting HEp-2 IFA. Results obtained were analyzed for accuracy with the intended and consensus response for three main categorical patterns (nuclear, cytoplasmic and mitotic), common patterns and ICAP report nomenclatures. The distributions of antibody titers of specimens were also compared. Results: Laboratories differed in the categorical patterns reported; 8 reporting all patterns, 3 reporting only nuclear patterns and 5 reporting nuclear patterns with various combinations of other patterns. For all participants, accuracy with the intended response for the categorical nuclear pattern was excellent at 99% [95% confidence interval (CI): 97-100%] compared to 78% [95% CI: 67-88%] for the cytoplasmic, and 93% [95% CI: 86%-100%] for mitotic patterns. The accuracy was 13% greater for the common nomenclature [87%, 95% CI: 82-90%] compared to the ICAP nomenclature [74%, 95% CI: 68-79%] for all participants. Participants reporting all three main categories demonstrated better performances compared to those reporting 2 or less categorical patterns. The average accuracies varied between participant groups, however, with the lowest and most variable performances for cytoplasmic pattern specimens. The reported titers for all specimens varied, with the least variability for nuclear patterns and most titer variability associated with cytoplasmic patterns. Conclusions: Our study demonstrates significant accuracy for all participants in identifying the categorical nuclear staining as well as traditional pattern assignments for nuclear patterns. However, there was less consistency in reporting cytoplasmic and mitotic patterns, with implications for assigning competencies and training for clinical laboratory personnel.

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A Multicenter Analysis of Subjectivity of Indirect Immunofluorescence Test in Antinuclear Antibody Screening

Cited by 6 publications

References 17 publications

A single-centre study on abnormal antinuclear antibodies in children caused by intravenous infusion of gamma globulin

A single-centre study on abnormal antinuclear antibodies in children caused by intravenous infusion of gamma globulin

The antinuclear antibody HEp-2 indirect immunofluorescence assay: a survey of laboratory performance, pattern recognition and interpretation

Reporting of Antinuclear Antibody HEp-2 Indirect Immunofluorescence Assay: A Performance Survey

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