DETERMINATION OF DQB1 ALLELES USING PCR AMPLIFICATION AND ALLELE-SPECIFIC PRIMERS

Molecular genotyping of HLA class II genes is commonly carried out usi ng polymerase chain reaction (PCR) in combination with sequence-specif ic oligotyping (PCR-SSO) or a combination of the PCR and restriction f ragment length polymorphism methods (PCR-RFLP). However, the identific ation of the DQB1 type by PCR-SSO and PCR-RFLP is very time-consuming which is disadvantageous for the typing of cadaveric organ donors. We have developed a DQB1 typing method using PCR in combination with alle le-specific amplification (PCR-ASA), which allows the identification o f the 17 most frequent alleles in one step using seven amplification m ixtures. PCR allele-specific amplification HLA-DQB1 typing is easy to perform, and the results are easy to interpret in routine clinical pra ctice. The PCR-ASA method is therefore better suited to DQB1 typing fo r organ transplantation than other methods.