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目的 分析人类 A、B、O3种基因的碱基序列变异 ,建立 PCR直接序列分析 ABO基因分型方法。方法 采用 PCR-直接测序技术分析人类 A转移酶基因 c DNA的 2 33～ 433与 6 6 0～ 788两个区域。结果 在 2 33～ 433区域存在 2 5 8th、2 97th两个碱基替换变异 ;在 6 6 0～ 788区域存在 70 0 th1个碱基替换变异。在 2 5 8th碱基位置 ,A、B基因为碱基“G”,O基因为碱基“A”。在 2 97th碱基位置 ,A基因为碱基“A”,B基因为碱基“G”,O基因存在 OA、OG两种亚型。在 70 0 th碱基位置 A、O基因为碱基“G”,B基因为碱基“A”。据此 ,对 2 33～ 433区域分析可确定 AA、AOA、AB、BB、BOG、OAOA、OGOG及 OAOG 8种基因型。对 6 6 0～ 788区域分析可进一步区分 AOG、BOA两种基因型。结论 PCR-直接序列分析为 ABO基因进一步分型提供了一种有效的新方法
OBJECTIVE: To analyze the nucleotide sequence variation of A, B and O genes in human and to establish ABO genotyping by PCR direct sequence analysis. Methods PCR-direct sequencing was used to analyze the two regions of human 33T3 433 and 6 60 0788 human cotransferase genes. The results showed that the 2 58th and the 2 97th two-base substitution mutations were located in the region of 33333 and the 70 ° th1-base substitution mutation existed in the region of 6 60 ~ 788. In the 25th base, the A and B genes are base “G” and the O gene is base “A”. In the 2 97th base, the A gene is the base “A”, the B gene is the base “G”, and the O gene has two subtypes OA and OG. At the 700th base position A, the O gene is the base “G” and the B gene is the base “A”. Based on this, 8 genotypes of AA, AOA, AB, BB, BOG, OAOA, OGOG and OAOG can be identified from the 2 33 ~ 433 region analysis. The 6 6 0 ~ 788 regional analysis can further distinguish AOG, BOA two genotypes. Conclusion PCR-direct sequence analysis provides an effective new method for further typing of ABO genes