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 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.
Fujiwara
K
K
Japanese Red Cross Central Blood Center, Tokyo, Japan.
Tokunaga
K
K
Isa
K
K
Miyamoto
M
M
Wang
L
L
Akaza
T
T
Tadokoroo
K
K
Shibata
Y
Y
Juji
T
T
D016824:Antigens, Human Platelet / Q000097:blood*; D001483:Base Sequence; D001782:Blood Donors; D004247:DNA / Q000032:analysis*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D012680:Sensitivity and Specificity; D012699:Serology / Q000379:methods
D016824:Antigens, Human Platelet; D004247:DNA