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

Ann Rheum Dis. 2001 Dec;60(12):1131-6. doi: 10.1136/ard.60.12.1131.

Abstract

Objective: To provide data on (a) the probability of detecting antinuclear antibodies (ANA) in a large and consecutive cohort of serum samples referred for ANA testing and (b) the probability of detecting more specific antinuclear reactivities (anti-DNA and anti-extractable nuclear antigens (anti-ENA)) in serum samples with a positive screening test (indirect immunofluorescence on HEp-2 cells).

Methods: Serum samples from 10 550 consecutive patients sent to the laboratory for ANA detection were analysed. In ANA positive serum samples (23.5% of referred serum samples), ANA were identified by indirect immunofluorescence on Crithidia, by immunodiffusion, and by line immunoassay. Because anti-SSA 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 positive serum samples: anti-dsDNA in 3.2%; anti-ENA (anti-SSA 10.5%, anti-SSB 6.7%, anti-RNP 2.7%, anti-Sm 1.8%, anti-Scl70 1.2%, anti-Jo-1 0.2%) in 15.8%, rRNP and anti-Cenp-B in respectively 0.5% and 4.0%. Multiple reactivities 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 detection of anti-SSA antibodies by line immunoassay was confirmed by additional assays.

Conclusions: The data from this analysis are useful in estimating the probabilities of detecting specific ANA. Line immunoassay was shown to be a sensitive test for the detection of anti-ENA antibodies.

MeSH terms

  • Antibodies, Antinuclear / blood*
  • Antibody Specificity
  • Antigens, Nuclear
  • Autoantigens / immunology
  • Biomarkers / blood
  • Cohort Studies
  • Connective Tissue Diseases / blood
  • Connective Tissue Diseases / diagnosis*
  • DNA-Binding Proteins / immunology
  • Fluorescent Antibody Technique, Indirect / methods
  • Humans
  • Immunoassay / methods
  • Immunodiffusion / methods
  • Nuclear Proteins / immunology

Substances

  • Antibodies, Antinuclear
  • Antigens, Nuclear
  • Autoantigens
  • Biomarkers
  • DNA-Binding Proteins
  • Nuclear Proteins