HLA-DQB1 and DQA1 matching by ambient temperature PCR-SSCP

We have developed a polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) protocol for rapid matching of DQA1 and DQB1 alleles. Electrophoresis can be performed at ambient temperature within the range 18-28 degrees C without continuous gel cooling. The method has been tested on 27 patient-potential bone marrow donor pairs for DQB1 and 31 pairs for DQA1. Bone marrow pairs were chosen to represent a broad range of common alleles based upon previous restriction fragment length polymorphism (RFLP) analysis type assignments. Samples were re-typed by PCR with sequence-specific primers (PCR-SSP) and the results compared to matching by PCR-SSCP analysis. There was a 100% correlation between PCR-SSP and PCR-SSCP analysis for DQB1, and a 97% correlation for DQA1 matching.