Abstract

In this study, we aimed to assess the prevalence of uncommon staining patterns found during testing for the presence of antinuclear antibodies (ANA) and to determine their association with certain antibodies and clinical diagnoses. Presence of ANA and the staining pattern was determined in 10955 samples using indirect immunofluorescence (IIF) on HEp-2 cells. ANA-positive samples were assessed for presence of 14 specific antibody types using a microbead based system. Demographic data (age, sex) and clinical diagnoses were collected from the referral documentation. Particular staining patterns were then compared with a representative comparison group comprised of samples with common staining patterns using these criteria. There were 22 patterns present in less than 3% of samples each and these were jointly present in 42.43% of ANA-positive samples. Specific antibodies were found in proportions similar to the comparison group (46.06%) and varied significantly between patterns. Likewise, there were significant differences in antibody distribution in particular patterns. Some patterns were associated with presence of rheumatic diseases or inflammatory arthropathies, while in others there was a concurrent diagnosis of liver disease, or a neoplastic process. Many of the uncommon IIF patterns have distinctive characteristics that warrant further investigation in order to determine their role in diagnosing various diseases, not limited only to the illnesses of the rheumatic spectrum. IIF on HEp-2 cells remains an irreplaceable method because of the diversity of ANA, only a number of which can be detected using other standardised methods.

Highlights

  • Ever since their discovery, antibodies directed against cellular antigens, commonly referred to as antinuclear antibodies (ANA), have been a significant tool in the diagnostic approach to patients with a suspicion of autoimmune illness [1,2]

  • The aim of this study is to assess the prevalence of unusual indirect immunofluorescence (IIF) patterns on HEp-2 cells in a setting of a tertiary facility with a high throughput of patients referred for the detection of ANA, to assess main demographic characteristics of the patients in whom they are observed, and to comprehensively analyse their potential associations with particular specific autoantibodies and clinical diagnoses

  • This study demonstrates why it is of paramount importance to continue screening for ANA using the proven manual method that is IIF microscopy on HEp-2 cells, a procedure that gains relevance only through adequate expertise and adherence to the latest recommendations

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Summary

Introduction

Antibodies directed against cellular antigens, commonly referred to as antinuclear antibodies (ANA), have been a significant tool in the diagnostic approach to patients with a suspicion of autoimmune illness [1,2]. Despite of recent advancements in developing commercial assays based on a predefined mixture of cellular antigens [12,13,14], the preferred method for the initial assessment of ANA is still indirect immunofluorescence (IIF) on HEp-2 cells [15] This method allows, provided that the competence of personnel performing the test is adequate, the detection of the presence of any autoantibody directed against cellular antigens, even of unknown significance or specificity, rather than a finite number of known common antigens [16,17]. The negative result of a confirmatory test does not exclude the existence of an unknown antibody type, and leaves both clinicians and laboratory personnel with a dilemma regarding the significance of a positive IIF test in these cases

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