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. 2013:2013:978202.
doi: 10.1155/2013/978202. Epub 2013 Nov 27.

Highlights on novel technologies for the detection of antibodies to Ro60, Ro52, and SS-B

M Infantino  1 , C Bentow  2 , A Seaman  2 , M Benucci  3 , F Atzeni  4 , P Sarzi-Puttini  4 , B Olivito  1 , F Meacci  1 , M Manfredi  1 , M Mahler  2
Affiliations

Highlights on novel technologies for the detection of antibodies to Ro60, Ro52, and SS-B

M Infantino et al. Clin Dev Immunol. 2013.

Abstract

Objective: We aimed to compare a chemiluminescent immunoassay (CIA, QUANTA Flash) on BIO-FLASH with a multiplex flow immunoassay (MFI) on BioPlex 2200 for the detection of antibodies to Ro60, Ro52, and SS-B.

Methods: The study included 241 samples, from patients suffering from systemic autoimmune diseases (n = 108) as well as disease controls (n = 133). All samples were tested for anti-Ro52, anti-Ro60, and anti-SS-B (La) antibodies on QUANTA Flash (INOVA Diagnostics, San Diego, USA) and BioPlex 2200 (Bio-Rad Laboratories Inc., Hercules, USA). Discrepant samples were tested by two independent methods: BlueDot/ANA and QUANTRIX Microarray (both D-tek, Belgium).

Results: The overall qualitative agreements were 95.4% (95% confidence interval, CI 92.0-97.7%) for anti-Ro52, 98.8% (95% CI 96.4-99.7%) for anti-Ro60, and 91.7% (95% CI 87.5-94.9%) for anti-SS-B antibodies. There were 34 discrepant samples among all assays (20 anti-SS-B, 11 anti-Ro52, 3 anti-Ro60). 30/33 of retested samples (by D-tek dot blot) agreed with the QUANTA Flash results. Similar findings were obtained with QUANTRIX Microarray kit.

Conclusion: QUANTA Flash and BioPlex 2200 show good qualitative agreement. The clinical performances were similar for anti-Ro52 and anti-Ro60 autoantibodies while differences were observed for anti-SS-B (La) antibodies.

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Figures

Figure 1
Figure 1
Comparison of QUANTA Flash with BioPlex 2200 as comparative method. Receiver operating characteristic (ROC) curves are shown in (a)–(c) and Spearman's correlation diagrams in (d)–(f). Note: A significant portion of positive samples are above the analytical measuring range of BioPlex which biases the Spearman rho values. PPA/NPA/TPA: Positive/Negative/Total % agreement. AUC: area under the curve.
Figure 2
Figure 2
Prevalence and titers of anti-Ro60, Ro52, and SS-B antibodies. Results for anti-Ro60 are shown in (a) and (b), for anti-Ro52 in (c) and (d), and for anti-SS-B antibodies in (e) and (f). AT: Atopic dermatitis, HI: healthy individuals, RA: rheumatoid arthritis, SED: suspected eye disease, SLE: systemic lupus erythematosus, SjS: Sjögren's syndrome, SSc: systemic sclerosis, UCTD: undifferentiated connective tissue disease.
Figure 3
Figure 3
Clinical comparative ROC analysis. Results of patients with systemic autoimmune rheumatic diseases were compared with controls. Cut-off values are indicated by arrows. For sensitivity and specificity, see Table 1. Results for anti-Ro60 (a), anti-Ro52 (b), and anti-SS-B antibodies (c) are shown.
Figure 4
Figure 4
Supervised cluster analysis of the results. Supervised centered cluster analysis according to disease cohort is shown. The dendrogram shows that the Ro60 assays are closely related, the Ro52 assays are somewhat related, and the SS-B assays show significant difference. AT: atopic dermatitis; UCTD: undifferentiated connective tissue disease, RA: rheumatoid arthritis, SSc: systemic sclerosis; SED: suspected eye disease, SLE: systemic lupus erythematosus; HI: healthy individuals; SARD: systemic autoimmune rheumatic disease.
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