Autoantibody profiling of patients with primary biliary cirrhosis using a multiplexed line-blot assay

Clin Chim Acta. 2015 Jan 1:438:135-8. doi: 10.1016/j.cca.2014.08.024. Epub 2014 Aug 27.

Abstract

Objective: To evaluate the autoantibody profile in patients with primary biliary cirrhosis (PBC) using a new multiplexed line-blot assay specifically designed for the diagnosis of autoimmune liver diseases.

Methods: Sera of 58 consecutive PBC patients and 191 disease controls (144 with autoimmune liver diseases other than PBC, and 67 with non-autoimmune chronic liver diseases) were tested by both the multiplexed line-blot Autoimmune Liver Disease Profile 2 (ALD2) and by IIF on HEp-2 cells and on rat kidney/liver/stomach tissues. ALD2 contains the following PBC-associated antigens: AMA-M2, natively purified from bovine heart; M2-E3, a recombinant fusion protein including the E2 subunits of PDC, BCOADC and OGDC; sp100, PML and gp210 recombinant proteins.

Results: With the ALD2 assay, a positive reaction to AMA-M2, M2-E3, sp100, PML and gp210 in PBC patients was observed in 77.6%, 84.5%, 34.5%, 15.1% and 18.9%, respectively, of the PBC sera. The overall sensitivity and specificity for PBC were 98.3% and 93.7%. Using IIF, positivity rates to AMA, and to antinuclear autoantibodies with membranous/rim-like and multiple nuclear dot patterns were 86.2%, 8.6% and 22.4%, respectively. The overall sensitivity and specificity for PBC of the IIF method were 86.2% and 97.9%, respectively.

Conclusions: The ALD2 line-blot showed a good diagnostic accuracy for PBC and a higher sensitivity than the IIF method to detect sp100 and gp210 autoantibodies.

Keywords: Anti-mitochondrial autoantibodies (AMA); Autoantibodies; Membranous/rim-like; Multiple nuclear dots (MND); gp210; sp100.

Publication types

  • Comparative Study

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Animals
  • Autoantibodies / blood*
  • Autoantigens / immunology*
  • Biological Assay
  • Biomarkers / blood*
  • Case-Control Studies
  • Cattle
  • Child
  • Child, Preschool
  • Female
  • Follow-Up Studies
  • Humans
  • Liver Cirrhosis, Biliary / blood
  • Liver Cirrhosis, Biliary / diagnosis*
  • Liver Cirrhosis, Biliary / immunology
  • Male
  • Middle Aged
  • Prognosis
  • Rats
  • Sensitivity and Specificity
  • Young Adult

Substances

  • Autoantibodies
  • Autoantigens
  • Biomarkers