Study of FUT2 gene in urinary sediment samples

ENSINCK A; RACCA L; GARCÍA BORRÁS S; YABER F; PROVENZAL O; COTORRUELO C; BIONDI C

Londres

Congreso; 25th Regional Congress of the International Society of Blood Transfusion; 2015

Backgraund: Histo-blood group ABH antigens are major alloantigens in humans. These antigens are widely distributed in human tissues and are synthesised through a a(1,2) fucosylatranferase (FUT2) which incorporates molecules of fucose in common type oligosaccharide precursor. The FUT2 gene has a significant polymorphism with typical ethnic specificity. The nonsense mutation 428G?A (Trp143?stop) is characteristic for the dominating nonsecretor allele (se428) in Europeans and ppearsin about 20% of the Caucasian population. These individuals are no secretors who fail to express soluble A, B, H, and Leb histo-boold group antigens in secretor individuals and secretor fluid because the absence of the Se (FUT2) gene. Aim: To study from urinary sediment samples, the allelic varieties of the FUT2 geneby a PCR reaction. Methods: Frozen saliva and urinary sediment samples from 136 unrelated of a population of Argentine were examined in this study. Appropriate informed consent was obtained from all subjects and all procedures were performed according to the ethical standards established by the University of Rosario. We determinated the secretor status in saliva with the hemagglutination inhibition technique using monoclonal antibodies anti-A, anti-B and lectin ulex europeaus. Agglutination of cells by antibody in tubes containing saliva samples indicates that the corresponding antigen is not present (non secretor status). Failure of known antibody to agglutinate indicator cells after incubation with saliva indicates that contains the corresponding antigen (secretor status). For the molecular studies the sediment urinary samples were centrifugated and the genomic DNA was extracted from the pellet by an enzymatic digestion method. The DNA samples were analyzed by ASA-PCR with specific primers for the G428A allele and for the wild type allele of the FUT2 gene. The PCR products (132 bp) were analyzed in 2% agarose gel containing ethidium bromide. Results: The results obtained by serological and molecular methods presented 100% of concordance. Both techniques indicated that the 78% of the nvestigate individuals were secretors. The G428A polymorphism had present in 9.3%, smaller value to report in the bibliography for the Caucasian population. Conclusions: This preliminary study demonstrates that allelic varieties of the FUT2gen can be investigated using urinary sediment samples which were obtained withnon-invasive methods. This method designed in our laboratory is reproducible androbust. The allelic varieties of the other non-secretor individuals different to theG428A might to correspond to other mutations.