Simultaneous genotyping of human platelet alloantigen-1 to 28bw systems by multiplex polymerase chain reaction sequence-based typing

Vox Sang. 2017 May;112(4):360-366. doi: 10.1111/vox.12507. Epub 2017 Mar 31.

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.

Keywords: Chinese population; PCR-SBT; genotype frequency; human platelet alloantigen; multiplex PCR.

MeSH terms

  • Antigens, Human Platelet / genetics*
  • Asian People / genetics
  • Female
  • Gene Frequency*
  • Genotyping Techniques / methods*
  • Humans
  • Male
  • Multiplex Polymerase Chain Reaction / methods*
  • Thrombocytopenia / diagnosis*
  • Thrombocytopenia / genetics

Substances

  • Antigens, Human Platelet