Polymorphism FUT2 gene: analysis in saliva samples

MORENO A; RACCA A; VALDEZ V; GARCÍA BORRÁS S; RACCA L; COTORRUELO C; BIONDI C

El Cairo - Egipto

Congreso; XIX Regional Congress. International Society of Blood Transfusion.; 2009

International Society of Blood Transfusion

Backgraund: The ABH and Lewis antigens on mucosa are synthesised through a a(1,2) fucosylatranferase (FUT2) which incorporates molecules of fucose in common type oligosaccharide precursor. The Se (FUT2) gene encoded a(1,2) fucosylatranferase activity. About 20% - 25% of Caucasian individuals are no secretors who fail to express soluble A,B,H, and Leb histo-boold group antigens in secretor organs and secretor fluid because the absence of the Se (FUT2) gene. A nonsense mutation (G428A) causes the nonsecretor phenotype in this population. Aim: to study from saliva samples, the allelic varieties of the FUT2 gene by a PCR reaction. Methods: Frozen saliva samples from 108 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 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 76 % of the investigate individuals were secretors. The G428A polymorphism had present in 8.5 %, smaller value to report in the bibliography for the Caucasian population. Conclusions: This preliminary study demonstrates that allelic varieties of the FUT2 gen can be investigated using saliva samples which were obtained with non-invasive methods. This method designed in our laboratory is reproducible and robust. The allelic varieties of the other non-secretor individuals different to the G428A might to correspond to other mutations.