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How should a district general hospital immunology service screen for anti‐nuclear antibodies? An ‘in‐the‐field’ audit

Anti‐nuclear antibody (ANA) testing assists in the diagnosis of several immune‐mediated disorders. The gold standard method for detection of these antibodies is by indirect immunofluorescence testing on human epidermoid laryngeal carcinoma (HEp‐2) cells. However, many laboratories test for these antibodies using solid‐phase assays such as enzyme‐linked immunosorbent assay (ELISA), which allows for higher throughput testing at reduced cost. In this study, we have audited the performance of a previously established ELISA assay to screen for ANA, making comparison with the gold standard HEp‐2 immunofluorescence test. A prospective and unselected sample of 89 consecutive ANA test requests by consultant rheumatologists were evaluated in parallel over a period of 10 months using both tests. ELISA and HEp‐2 screening assays yielded 40 (45%) and 72 (81%) positive test results, respectively, demonstrating lack of concordance between test methods. Using standard and clinical samples, it was demonstrated that the ELISA method did not detect several ANA with nucleolar, homogeneous and speckled immunofluorescence patterns. None of these ELISANEG HEp‐2POS ANA were reactive with a panel of six extractable nuclear antigens or with double‐stranded DNA. Nonetheless, 13 of these samples (15%) originated from patients with recognized ANA‐associated disease (n = 7) or Raynaud's phenomenon (n = 6). We conclude that ELISA screening may fail to detect clinically relevant ANA that lack defined specificity for antigen.