ICARE improves antinuclear antibody detection by overcoming the barriers preventing accreditation

Antinuclear antibodies (ANA) are useful biomarkers for the diagnosis and the monitoring of rheumatic diseases. The American College of Rheumatology has stated that indirect immunofluorescence (IIF) analysis remains the gold standard for ANA screening. However, IIF is time consuming, subjective, not fully standardized and presents several issues for accreditation which is the process leading to ISO 15189 certification for medical laboratories. We propose an innovative tool for accreditation by using the quantitative evaluation of the automated image capture and analysis "ICARE" (Immunofluorescence for Computed Antinuclear antibody Rational Evaluation). We established the optimal screening dilution (1:160) and a fluorescence index (FI) cutoff for ICARE on a cohort of 91 healthy blood donors. Then, we evaluated performance of ICARE on a routine cohort of 236 patients. Precision parameters of ANA detection by IIF were evaluated according to ISO 15189. ICARE showed an excellent concordance with visual evaluation (88%, Kappa = 0.76) and significantly discriminated between weak to moderate (1:160–1:320 titers) and high (> 1:320 titers) ANA levels. A significant correlation was found between FI and ANA titers (Spearman's ρ = 0.67; P < 0.0001). Using ICARE, we reported precision parameters such as repeatability (CV < 13.8%) and reproducibility (CV < 13.1%) as well as absence of inter-sample contamination for ANA detection by IIF according to ISO 15189 standards. ICARE offers a precious help for the accreditation of IIF qualitative methods. This innovative quantitative approach is in adequacy with the process of continuous improvement of the quality of clinical laboratories.