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目的构建1个简单、有效,用于ABO基因分型的AS-PCR体系,及基于AS-PCR体系的PCR测序方法,并将此方案用于1个B亚型家系分析。方法参照ABO血液基因型cDNA的261,297,467和803位点,通过设计针对这些位点的AS-PCR引物构建可用于ABO血液基因型分型的AS-PCR体系,并将该AS-PCR体系应用于1个B亚型家系进行分析,并依据其分析结论设计PCR测序方法,以进行单体型分型。结果该家系成员母亲和两名子女的基因型分别为O01Bw11、O01Bw11和O02Bw11。结论 AS-PCR体系和PCR测序相结合的方案,操作简单、耗时短、结果准确,可用于ABO亚型的基因型分型。
OBJECTIVE: To construct a simple and effective AS-PCR system for ABO genotyping and PCR-based sequencing method based on AS-PCR system, and to use this protocol for analysis of one B subtype family. Methods Based on the 261,297,467 and 803 loci of ABO blood genotype cDNAs, AS-PCR primers that could be used to genotype ABO blood were constructed by designing AS-PCR primers targeting these loci. The AS-PCR system was applied to 1 A subtype B family analysis, and based on its analysis of the design of PCR sequencing method for haplotyping. Results The genotypes of members of the pedigree and two children were O01Bw11, O01Bw11 and O02Bw11, respectively. Conclusion The combination of AS-PCR system and PCR-based sequencing is simple, time-consuming and accurate. It can be used to genotype ABO subtypes.