DETERMINATION OF DRB ALLELES USING PCR AMPLIFICATION AND ALLELE-SPECIFIC PRIMERS

HLA class-II allelic diversity is commonly defined using polymerase ch ain reaction (PCR) in combination with sequence-specific oligotyping ( PCR-SSO) or the combination of PCR and restriction fragment length pol ymorphism methods (PCR-RFLP). Nevertheless, the identification of the DRB polymorphism by PCR-SSO is a time-consuming procedure and the PCR- RFLP is cumbersome. A rapid technique which allows a precise and exten sive HLA-DRB typing is required, particularly in order to study the ro le of class-II matching in organ transplantation. A DRB typing method based on the detection and length of PCR products amplified using comb ination of allele specific primers has been developed. Thirty-four DRB alleles (29 DRB1, 4DRB3, 1DRB4) can be detected using 29 primers dist ributed into 19 amplification mixtures.