Detection and identification of antinuclear antibodies (ANA) in a large and consecutive cohort of serum samples referred for ANA testing

Objective-To provide data on (a) the probability of detecting antinuclear a ntibodies (ANA) in a large and consecutive cohort of serum samples referred for ANA testing and (b) the probability of detecting more specific antinuc lear reactivities (anti-DNA and anti-extractable nuclear antigens (and-ENA) ) in serum samples with a positive screening test (indirect immunofluoresce nce on HEp-2 cells). Methods-Serum samples from 10 550 consecutive patients sent to the laborato ry for ANA detection were analysed. In ANA positive serum samples (23.5% of referred serum samples), ANA were identified by indirect immunofluorescenc e on Crithidia, by immunodiffusion, and by line immunoassay. Because anti-S SA antibodies were the most frequently identified ANA, sensitively detected by line immunoassay, additional immunoassays were developed to confirm the specificity of the line immunoassay result. Results-At least one fine reactivity could be identified in 21.1% of ANA po sitive serum samples: anti-dsDNA in 3.2%; anti-ENA (anti-SSA 10.5%, anti-SS B 6.7%, anti-RNP 2.7%, anti-Sm. 1.8%, anti-Scl70 1.2%, anti-Jo-1 0.2%) in 1 5.8%, rRNP and anti-Cenp-B in respectively 0.5% and 4.0%. Multiple reactivi ties were found in 7.9%. For anti-ENA antibodies, line immunoassay was more sensitive than immunodiffusion (15.4% v 7.7%; p <0.0001). The sensitive de tection of anti-SSA antibodies by line immunoassay was confirmed by additio nal assays. Conclusions-The data from this analysis are useful in estimating the probab ilities of detecting specific ANA. Line immunoassay was shown to be a sensi tive test for the detection of anti-ENA antibodies.