PCR with sequence-specific primer-based simultaneous genotyping of human platelet antigen-1 to -13w

Autor: Hui-Yu Hu, Ying-Ju Chen, Jau-Yi Lyou, Jeong-Shi Lin, Cheng-Hwai Tzeng
Rok vydání: 2002
Předmět:
Zdroj: Transfusion. 42:1089-1095
ISSN: 1537-2995
0041-1132
Popis: BACKGROUND : Accurate human platelet antigen (HPA) typing is important for patients with diagnosis of alloimmune thrombocytopenic syndromes and provision of HPA-matched blood components for these patients. STUDY DESIGN AND METHODS : Thirteen sequence-specific primers (SSPs) designed on the basis of known published polymorphisms for HPA-1 to HPA-13w, respectively, were employed for simultaneous HPA genotyping. All PCR amplifications were carried out with identical cycling conditions in 96-well plates containing primer mixtures. A total of 300 blood samples from unrelated volunteer donors in Taiwan were included in the study. RESULTS : All primers had specific amplification products. The typing results were available within 4 hours each time for up to four blood samples tested. Among the 13 HPAs, HPA-3 had the greatest heterozygosity with a gene frequency of 0.3267, 0.4967, and 0.1767 for HPA-3a/HPA-3a, HPA-3a/HPA-3b, and HPA-3b/HPA3-b, respectively. For the remaining 12 HPAs, the predominance of a/a homozygosity was noted for HPA-1, -2, -4, -5, and -6, with a frequency ranging from 0.9200 to 0.9967. The frequency of a/a homozygosity was 1.0000 for HPA-7w to -13w, except for HPA-10w, for which one case was observed to be HPA-10aw/HPA-10bw heterozygous. Excluding HPA-3, b/b homozygosity was noted in only one case ( HPA-6b/HPA-6b ). The prevalence rates of HPA-1 to -13w in this study were consistent with previous reports using different methods. CONCLUSION : An extended, streamlined PCR-SSP protocol for simultaneous genotyping of HPA-1 to HPA-13w was established. This allows fast and reliable diagnosis of alloimmune thrombocytopenia, and is readily applicable to large-scale genetic population studies.
Databáze: OpenAIRE
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