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  • Title: DNA-based typing of human platelet antigen systems by polymerase chain reaction-single-strand conformation polymorphism method.
    Author: Fujiwara K, Tokunaga K, Isa K, Miyamoto M, Wang L, Akaza T, Tadokoroo K, Shibata Y, Juji T.
    Journal: Vox Sang; 1995; 69(4):347-51. PubMed ID: 8751305.
    Abstract:
    Polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) method has been established to discriminate genotypes for the human platelet antigen (HPA) systems HPA-1, HPA-2, HPA-3, HPA-4, and HPA-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 silver staining. The results obtained by the PCR-SSCP method were in good agreement with those of the allotypes determined by serological typing. Furthermore, the results agreed with those obtained by other DNA-based typing methods such as PCR-allele-specific restriction enzyme analysis and PCR-sequence-specific primer. These results indicate that PCR-SSCP is a simple and sensitive method for determining HPA genotypes and identifying unknown polymorphisms.
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