基因测序方法在ABO血型鉴定中的应用  

Application of gene sequencing method in ABO blood group identification

作　　者：刘建成 邵峰 步晓筠 杨洁 毛小尹 海静 LIU Jiancheng;SHAO Feng;BU Xiaoyun;YANG Jie;MAO Xiaoyin;HAI Jing(Blood Group Reference Laboratory,Ningxia Blood Center,Yinchuan,Ningxia 750000,China)

机构地区：[1]宁夏血液中心血型参比实验室,银川750000

出　　处：《重庆医学》2024年第11期1690-1694,共5页Chongqing medicine

基　　金：宁夏卫生健康系统科研课题项目(2022-NWKY-040)。

摘　　要：目的 建立一种ABO血型基因测序方法,在DNA水平对突变位点进行分析,以准确鉴定ABO血型。方法 选取20份血液标本,其中18份血清学鉴定为ABO正常血型,2份为ABO亚型。利用序列特异性引物PCR(PCR-SSP)方法,对ABO血型基因第6、7外显子进行扩增,然后进行PCR直接测序分析基因序列,与ABO参考序列进行比对分析鉴定ABO血型。结果 20份血液标本基因测序鉴定结果与血清学结果一致。2份亚型标本中1份基因型为BA.02/O.01,表型为B(A)亚型,其第7外显子700位发生了C>G突变,导致氨基酸翻译时第234位脯氨酸变为丙氨酸。另1份基因型为AW.37/B.01,表型为AxB亚型,其第7外显子940位发生A>G突变,导致蛋白质翻译时第314位赖氨酸变为谷氨酸。结论 建立了一种适合实验室的ABO血型基因测序方法,能够准确鉴定ABO血型。Objective To establish a gene sequencing method for ABO blood group,to analyze the mutation sites at the DNA level in order to accurately identify ABO blood group.Methods Twenty blood samples were selected,in which 18 samples were ABO normal blood group and 2 samples were the ABO subtype.Exons 6 and 7 of ABO blood group gene were amplified by sequence-specific primer PCR(PCR-SSP),and then the gene sequence was directly sequenced and analyzed by PCR,and the ABO blood group was identified by comparing with the ABO reference sequence.Results The gene sequencing results of 20 blood samples were consistent with the serological results.In 2 subtype samples,the genotype in 1 sample was BA.02/O.01 and its phenotype was B(A)subtype.C>G mutation occurred at position 700 of the 7th exon,which resulted in proline changing to alanine at position 234 during amino acid translation.The genotype of the other sample was AW.37/B.01 and the phenotype was AxB subtype.The position 940 of the 7th exon mutates from adenine to guanine,resulting in the mutation of lysine changing to glutamic acid at position 314 during protein translation.Conclusion A method of ABO blood group gene sequencing suitable for laboratory is established,which could accurately identify ABO blood group.

关 键 词：ABO血型 基因测序 ABO亚型 ABO基因 外显子 

分 类 号：R457.11[医药卫生—治疗学]