Assessment of antinuclear antibodies by indirect immunofluorescence assay: report from a survey by the American Association of Medical Laboratory Immunologists

Abstract

BackgroundThe indirect immunofluorescence assay (IFA) using HEp-2 cell substrates is the preferred method by some for detecting antinuclear antibodies (ANA) as it demonstrates a number of characteristic staining patterns that reflect the cellular components bound as well as semi-quantitative results. Lack of harmonized nomenclature for HEp-2 IFA patterns, subjectivity in interpretation and variability in the number of patterns reported by different laboratories pose significant harmonization challenges. The main objectives of this study were to assess current practice in laboratory assessment of HEp-2 IFA, identify gaps and define strategies to improve reading, interpretation and reporting.MethodsWe developed and administered a 24-item survey based on four domains: educational and professional background of participants, current practice of HEp-2 IFA testing and training, gap assessment and the perceived value of International Consensus on Antinuclear Antibody Patterns (ICAP) and other factors in HEp-2 IFA assessment. The Association of Medical Laboratory Immunologists (AMLI) and American Society for Clinical Pathology administered the survey from April 1 to June 30, 2018, to members involved in ANA testing. This report summarizes the survey results and discussion from a dry workshop held during the 2019 AMLI annual meeting.ResultsOne hundred and seventy-nine (n = 179) responses were obtained where a significant number were clinical laboratory scientists (46%), laboratory directors (24%), supervisors (13%) or others (17%). A majority of respondents agreed on the need to standardize nomenclature and reporting of HEp-2 IFA results. About 55% were aware of the ICAP initiative; however, among those aware, a significant majority thought its guidance on HEp-2 IFA nomenclature and reporting is of value to clinical laboratories. To improve ICAP awareness and further enhance HEp-2 IFA assessment, increased collaboration between ICAP and the clinical laboratory community was suggested with emphasis on education and availability of reference materials.ConclusionsBased on these suggestions, future efforts to optimize HEp-2 IFA reading, interpretation and reporting would benefit from more hands-on training of laboratory personnel as well as continuous collaboration between professional organizations, in vitro diagnostic manufacturers and clinical laboratories.

Current laboratory and clinical practices in reporting and interpreting anti-nuclear antibody indirect immunofluorescence (ANA IIF) patterns: results of an international survey

“…The familiarity with ANA IIF patterns is mainly clear for the nuclear competent patterns (with the exception of DFS) and the cytoplasmic reticular/mitochondria-like antibody (AC-21) pattern, findings which are in concordance to the results of a similar recent survey performed by the American association of Medical Laboratory Immunologists [ 24 ]. Clinicians were less familiar with the cytoplasmic reticular pattern than laboratory professionals, which might be related to the fact that the survey was mainly directed to rheumatologists (and not hepato-gastroenterologists).…”

Current laboratory and clinical practices in reporting and interpreting anti-nuclear antibody indirect immunofluorescence (ANA IIF) patterns: results of an international survey. 

“…The familiarity with ANA IIF patterns is mainly clear for the nuclear competent patterns (with the exception of DFS) and the cytoplasmic reticular/mitochondria-like antibody (AC-21) pattern, ndings which are in concordance to the results of a similar recent survey performed by the American association of Medical Laboratory Immunologists (24). Clinicians were less familiar with the cytoplasmic reticular pattern than laboratory professionals, which might be related to the fact that the survey was mainly directed to rheumatologists (and not hepatogastroenterologists).…”

Standardization and Quality Assessment Under the Perspective of Automated Computer-Assisted HEp-2 Immunofluorescence Assay Systems

“…It is necessary that the biomedical industry produces a further effort aimed both at expanding the spectrum of patterns that can be identified (for example the dense fine speckled) consistently with those classified by ICAP, and at the recognition of mixed patterns (35,79). The implementation of the ICAP nomenclature, despite being already widespread, is believed to be only a first step towards the common goal of harmonizing the interpretation of HEp-2 IIF tests.…”