DNA-BASED TYPING OF HUMAN PLATELET ANTIGEN SYSTEMS BY POLYMERASE CHAIN REACTION-SINGLE-STRAND CONFORMATION POLYMORPHISM METHOD

Polymerase chain reaction-single-strand conformation polymorphism (PCR -SSCP) method has been established to discriminate genotypes for the h uman platelet antigen (HPA) systems HPA-1, HPA-2, HPA-3, HPA-4, and HP A-5. Gene fragments which contain polymorphic sequences corresponding to the HPA-1, HPA-2 HPA-3, HPA-4, and HPA-5 systems were PCR-amplified with specific primers. The amplified DNA was denatured and subjected to non-denaturing polyacrylamide gel electrophoresis followed by silve r staining. The results obtained by the PCR-SSCP method were in good a greement with those of the allotypes determined by serological typing. Furthermore, the results agreed with those obtained by other DNA-base d typing methods such as PCR-allele-specific restriction enzyme analys is and PCR-sequence-specific primer. These results indicate that PCR-S SCP is a simple and sensitive method for determining HPA genotypes and identifying unknown polymorphisms.