X Hong1,2,3, S Chen1,2,3, Y Ying1,2,3, Y Liu1,2,3, X Xu1,2,3, J He1,2,3, F Zhu1,2,3. 1. Blood Center of Zhejiang Province, Hangzhou, Zhejiang, China. 2. Key Laboratory of Blood Safety Research, Ministry of Health, Hangzhou, Zhejiang, China. 3. Zhejiang Provincial Key Laboratory of Blood Safety Research, Hangzhou, Zhejiang, China.
Abstract
BACKGROUND AND OBJECTIVES: Human platelet alloantigen (HPA) genotyping is important for the diagnosis and prevention the alloimmune platelet disorders. In this study, a simultaneous genotyping method for HPA-1 to -28bw systems was established using multiplex PCR-SBT and the frequencies of genotypes and alleles of HPA-1 to -28bw systems in the Zhejiang Han population were analysed. MATERIALS AND METHODS: The specific primers were designed according to the nucleotide sequences of HPA-1 to 28bw systems which are located in ITGB3, GP1BA, ITGA2B, ITGA2, GP1BB and CD109, respectively. The multiplex PCR amplification systems were used, and then, the amplicons were purified and sequenced. A total of 335 healthy volunteer blood donors were detected. RESULTS: The genotypes of ten reference samples from Platelet Immunology Workshop of ISBT were in concordance with the known genotypes. Among the 28 HPA systems, HPA a and b alleles were found in HPA-1 to 6w, HPA-15 and HPA-21w systems in the Chinese Han population, while only HPA aa genotype was detected in the other HPA systems. The frequencies of HPA-1a and HPA-1b were 0·993 and 0·007, with 0·943 and 0·057 for HPA-2a and HPA-2b, 0·527 and 0·473 for HPA-3a and HPA-3b, 0·997 and 0·003 for HPA-4a and HPA-4b, 0·991 and 0·009 for HPA-5a and HPA-5b, 0·980 and 0·020 for HPA-6wa and HPA-6wb, 0·508 and 0·492 for HPA-15a and HPA-15b and 0·994 and 0·006 for HPA-21wa and HPA-21wb. CONCLUSIONS: One multiplex PCR-SBT method for HPAs was established and the data of the study could help to prevent and treat for alloimmune thrombocytopenia.
BACKGROUND AND OBJECTIVES:Human platelet alloantigen (HPA) genotyping is important for the diagnosis and prevention the alloimmune platelet disorders. In this study, a simultaneous genotyping method for HPA-1 to -28bw systems was established using multiplex PCR-SBT and the frequencies of genotypes and alleles of HPA-1 to -28bw systems in the Zhejiang Han population were analysed. MATERIALS AND METHODS: The specific primers were designed according to the nucleotide sequences of HPA-1 to 28bw systems which are located in ITGB3, GP1BA, ITGA2B, ITGA2, GP1BB and CD109, respectively. The multiplex PCR amplification systems were used, and then, the amplicons were purified and sequenced. A total of 335 healthy volunteer blood donors were detected. RESULTS: The genotypes of ten reference samples from Platelet Immunology Workshop of ISBT were in concordance with the known genotypes. Among the 28 HPA systems, HPA a and b alleles were found in HPA-1 to 6w, HPA-15 and HPA-21w systems in the Chinese Han population, while only HPA aa genotype was detected in the other HPA systems. The frequencies of HPA-1a and HPA-1b were 0·993 and 0·007, with 0·943 and 0·057 for HPA-2a and HPA-2b, 0·527 and 0·473 for HPA-3a and HPA-3b, 0·997 and 0·003 for HPA-4a and HPA-4b, 0·991 and 0·009 for HPA-5a and HPA-5b, 0·980 and 0·020 for HPA-6wa and HPA-6wb, 0·508 and 0·492 for HPA-15a and HPA-15b and 0·994 and 0·006 for HPA-21wa and HPA-21wb. CONCLUSIONS: One multiplex PCR-SBT method for HPAs was established and the data of the study could help to prevent and treat for alloimmune thrombocytopenia.