HLA-DQB1 AND DQA1 MATCHING BY AMBIENT-TEMPERATURE PCR-SSCP

We have developed a polymerase chain reaction-single strand conformati on polymorphism (PCR-SSCP) protocol for rapid matching of DQA1 and DQB 1 alleles. Electrophoresis can be performed at ambient temperature wit hin the range 18-28 degrees C without continuous gel cooling. The meth od has been tested on 27 patient-potential bone marrow donor pairs for DQB1 and 31 pairs for DQA1. Bone marrow pairs were chosen to represen t a broad range of common alleles based upon previous restriction frag ment length polymorphism (RFLP) analysis type assignments. Samples wer e re-typed by PCR with sequence-specific primers (PCR-SSP) and the res ults compared to matching by PCR-SSCP analysis. There was a 100% corre lation between PCR-SSP and PCR-SSCP analysis for DQB1, and a 97% corre lation for DQA1 matching.