MOLECULAR HLA-B27 DETECTION IN PATIENTS WITH SPONDYLOARTHROPATHIES

SALVARREDI, LEONARDO; LACOSTE, MARÍA GABRIELA; GUZMÁN, ANA MARÍA.; DI GENARO, MARÍA SILVIA

Huerta Grande, Córdoba

Congreso; I Reunión Conjunta de Sociedades de Biología de la República Argentina; 2007

Asociación de Biología de Tucumán, Sociedad Argentina de Biología, Sociedades de Biología de Cuyo, Rosario y Córdoba

The human leukocyte antigen (HLA) molecules are associated in many cases with autoimmune diseases. HLA-B27 is strongly associated with spondyloarthropathies (SpA) in which HLA-B27 is a diagnostic and prognostic marker. Therefore, HLA-B27 determination results of clinical interest in patients with rheumatologicdiseases. The objective was to optimize an assay for HLA-B27 detection by molecular methods (PCR and real time- PCR) to be used in blood and synovial fluids (SF) of SpA patients. Two DNA salting out extraction methods were compared in blood and SFsamples. A conventional salting out method and an optimized method (with NaClO4) were used in 40 and 80 blood samples, respectively. The purity grade (DO 260/280) and the integrity were evaluated. The HLA-B27 genotypic determination was performedby PCR-SSP in clinical samples and in 100 aleatory selected blood samples from the Public San Luis Hospital. Co-amplification of HLA-B27 with β-globin gene as internal control was perfomed. DNA purity (DO 260/280) and integrity was higher in samples extracted with NaClO4 method than those extracted with the conventional method. One of 100 blood samples (1%) resulted HLAB27 positive. A SF sample from a patient resulted also HLA-B27 positive. This work contributed to optimize the molecular HLAB27 detection for application in patients with arthropathies.