Analytical and clinical performance of different platforms simultaneously detecting 15 antinuclear antibodies

Abstract

Background: Antinuclear antibodies (ANAs) are invaluable biomarkers for the diagnosis of autoimmune diseases (AIDs). This study aims to compare the performances of line immunoassay (LIA), multiplex bead-based flow fluorescent immunoassay (MBFFI), and magnetic bar code immunofluorescence assay (MBC-IF) to detect ANA-Profile-15S. Methods: In total, 184 samples from AID patients and 50 healthy controls (HCs) were collected. Fifteen ANAs (anti-dsDNA, nucleosome, histone, Sm, PCNA, ribosomal-P, SS-A/Ro52, SS-A/Ro60, SS-B/La, centromere B [CENP-B], Scl-70, U1-snRNP, AMA-M2, Jo-1, and Pm/Scl) were subjected to parallel detection by the LIA, MBFFI, and MBC-IF. The consistency between assays was analyzed. The discrepant results were further examined by chemiluminescent immunoassay (CLIA). Results: Anti-SS-A/Ro52 and SS-A/Ro60 autoantibodies were the most common autoantibodies in ANA positive-profiles, and were detected with equal efficiency by the LIA, MBFFI, and MBC-IF (p = 0.101 and p = 0.732, respectively). The three assays showed excellent agreement (consistency range: 66.5%-97.5%), and total consistency was 85.8%. The MBFFI and MBC-IF assays were in good agreement in terms of ANA-Profile-15S determination; the kappa coefficient ranged from 0.59 to 0.95, except for the PCNA and PM-Scl. Of the 262 re-assessed divergent results, 124 (47.33%) were positive on CLIA; the various autoantibodies exhibited variable patterns. More importantly, the ANA-Profile-15S results of the MBFFI and MBC-IF accurately identified patients with AID; the area under the curves ranged from 0.642 to 0.919. Conclusions: The novel MBFFI and MBC-IF assay performed well in detecting ANA-Profile-15S. The application of MBFFI and MBC-IF play important roles in laboratory diagnosis of AIDs.

Ro60 - Roles in RNA Processing, Inflammation, and Rheumatic Autoimmune Diseases

“…However, sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy of these diagnostic approaches may vary, and combinations of methods are recommended in clinical settings [89]. Using sera specimens of 184 autoimmune disease patients and 50 controls, Qin et al evaluated the analytical and clinical performance of line immunoassay, multiplex beadbased flow fluorescent immunoassay, and magnetic bar code immunoassay in detection of antinuclear antibodies, including those targeting Ro60 and Ro52; they found that the three assays showed good agreement [90]. Diagnostic accuracy is an important parameter in autoantibody detection, requiring consistent refinement of validation assays.…”