[Study on the simultaneous genotyping of human platelet antigens of 1, 2, 3, 4, 5, 6 system by PCR-SSP and its applications].
To set up the simultaneous genotyping of human platelet antigens of 1,2,3,4,5,6 system by PCR-SSP assay and use the genotyping method for the study of platelet antigens. In this study, 18 sequence-specific primers were designed and synthesized. The annealing temperature for all sequence-specific primer pair, the concentration of each primer pair and the concentration of Mg2+ were adjusted to the optimum so that HPA-1 to 6 systems could be amplified simultaneously under the same PCR cycling parameters. The electrophoresis of PCR products was conducted simultaneously on the same agarose gel. Control DNA samples that genotypes known were used to confirm the sensitivity and specificity of each sequence-specific primer. 15 coded samples (including 2 blood samples and 13 DNA samples) distributed by 10TH Platelet Genotyping and Serology Workshop of the International Society of Blood Transfusion (ISBT) were typed for HPA-1 to 6 systems by this method. A concordance rate of 100 percent was observed between the results of control DNA samples typed by our PCR-SSP assay and the data of known specificity of control DNA. The results of 15 coded samples tested by our method agreed well with the results provided by ISBT report.
Deng
Zhi-Hui
ZH
ShenZhen Institute of Transfusion Medicine, ShenZhen, Guangdong Province 518 035, China. zhihui_deng@yahoo.com.cn
Wu
Guo-Guang
GG
Li
Da-Cheng
DC
D016824:Antigens, Human Platelet / Q000145:classification / Q000235:genetics*; D004247:DNA / Q000032:analysis* / Q000235:genetics; D017931:DNA Primers; D004586:Electrophoresis / Q000379:methods; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods
D016824:Antigens, Human Platelet; D017931:DNA Primers; D004247:DNA