RAPID HLA-DQB TYPING BY 8 POLYMERASE CHAIN-REACTION AMPLIFICATIONS WITH SEQUENCE-SPECIFIC PRIMERS (PCR-SSP)

Molecular genotyping of HLA class II genes using group-specific DNA am plification by the PCR followed by probing with (PCR-SSO) probes is to o time consuming for the typing of cadaveric organ donors. Recently, a mplification of DNA using PCR-SSP has proved a reliable and rapid meth od for typing HLA-DRB1 genes. PCR-SSP takes 2 hours to perform and is therefore suitable for the genotyping of cadaveric donors. We have des igned a set of primers that in eight PCR reactions will positively ide ntify the HLA-DQB1 alleles corresponding to the serologically defined series HLA-DQ2, DQ4, DQ5, DQ6, DQ7, DQ8, and DQ9. Presently, 30 homozy gous cell lines and 138 individuals have been typed by the DQB1 PCR-SS P technique and compared with a combination of serology and RFLP with 100% concordance. No false-negative or false-positive amplifications w ere recorded. All combinations of DQB1 can be readily identified. DQB1 PCR-SSP can take as little as 130 minutes from start to finish, inclu ding DNA preparation.