Detection of serum M2 anti-mitochondrial antibodies by enzyme-linked immunosorbent assay is potentially less specific than by immunofluorescence

Ann Clin Biochem. 2002 May;39(Pt 3):304-7. doi: 10.1258/0004563021902008.

Abstract

Aim: To compare the predictive values of enzyme-linked immunosorbent assays (ELISAs) and indirect immunofluorescence (IIF) techniques for the detection of M2 anti-mitochondrial antibodies.

Methods: Commercial ELISAs are widely available for the detection of anti-mitochondrial antibody subtypes in primary biliary cirrhosis (PBC). We compared the results from two ELISAs (one recombinant, one purified antigen) with those from two IIF methods in a well-defined cohort of PBC patients and in patients with systemic lupus erythematosus, Sjögren's syndrome, sicca syndrome, systemic sclerosis, rheumatoid arthritis and blood donor controls.

Results: There was good correlation between a rodent substrate IIF and ELISA A (r=0.9134), but poor correlation with ELISA B (r=0.5999), which produced many false-positive results in the control population. We show that rodent IIF alone or human epithelial cell (HEp-2000) screening with confirmation by ELISA produce similar predictive values for PBC and require lesser degrees of skilled interpretation of IIF patterns.

Conclusions: We conclude that the specificities of IIF are greater than the ELISA methods (99% versus 85-97%), although the ELISAs are slightly more sensitive in biopsy-proven PBC. Careful in-house validation of all new ELISA technologies is mandatory for good laboratory practice, but IIF in experienced hands remains an effective and specific assay.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Animals
  • Antibody Specificity
  • Autoantibodies / blood*
  • Autoantibodies / immunology*
  • Autoimmune Diseases / diagnosis
  • Autoimmune Diseases / immunology
  • Cell Line
  • Enzyme-Linked Immunosorbent Assay / methods
  • Enzyme-Linked Immunosorbent Assay / standards*
  • False Positive Reactions
  • Female
  • Fluorescent Antibody Technique, Indirect / methods
  • Fluorescent Antibody Technique, Indirect / standards*
  • Humans
  • Liver Cirrhosis, Biliary / diagnosis
  • Liver Cirrhosis, Biliary / immunology
  • Male
  • Mitochondria / immunology*
  • Predictive Value of Tests
  • Rodentia
  • Sensitivity and Specificity

Substances

  • Autoantibodies