Jr(a-)稀有血型ABCG2^(*) 376T等位基因高分辨熔解曲线检测方法的建立及分布频率研究  

The establishment of a high-resolution melting curve genotyping method for ABCG2^(*) 376T allele of Jr(a-) and its distribution frequency study

作　　者：王贞[1] 贾双双[1] 廖志坚[1] 莫春妍[1] 魏玲[1] 张润青[1] 罗广平[1] 姬艳丽[1] WANG Zhen;JIA Shuangshuang;LIAO Zhijian;MO Chunyan;WEI Ling;ZHANG Runqing;LUO Guangpin;JI Yanli(Institute of Clinical Transfusion,Guangzhou Blood Center,Guangzhou 510095,China)

机构地区：[1]广州血液中心广州市血液安全重点实验室,广东广州510091

出　　处：《中国输血杂志》2022年第7期690-693,共4页Chinese Journal of Blood Transfusion

基　　金：广东省医学科学技术研究基金项目(编号:B2021225);广州市医学重点学科(2021-2023年)。

摘　　要：目的 建立针对Jr(a-)稀有血型ABCG2^(*)376T等位基因的高通量分子检测方法,并将其应用于中国人群该等位基因的分布频率研究。方法 设计针对ABCG2基因376位点的特异性引物,利用前期已获得的携带ABCG2^(*)376T等位基因的纯合子和杂合子标本作为阳性对照,不携带该突变的野生型标本作为阴性对照,建立该等位基因的高分辨熔解曲线(HRM)检测方法。应用建立的方法对广州地区献血人群的DNA标本进行筛查,对筛查出携带ABCG2^(*)376T等位基因的标本进行测序确证,以验证HRM方法的准确性。结果 成功建立可检测ABCG2^(*)376T等位基因,并可同时对纯合子及杂合子进行准确分型的HRM方法,在1 560例广州地区献血者中筛查出携带杂合型等位基因个体15例,尚未筛查出携带纯合型等位基因的个体。结论 HRM方法可用于准确筛查ABCG2^(*)376T等位基因并进行分型,该等位基因在中国人群的分布频率约为0.48%(15/3120)。Objective To establish a high-throughput detection method for ABCG2^(*)376T allele of Jr(a-),and apply it to the study of the frequency of this allele in the Chinese population.Methods The specific primers were designed and synthesized,the sample carrying homozygous ABCG2^(*)376T alleles,obtained in the previous study,was used as the homozygous positive control,and the sample carrying heterozygous allele as the heterozygous positive control.The wild-type sample was used as a negative control,and a high-resolution melting curve(HRM) method for detecting this allele was established.The established method was used to screen DNA samples from blood donors in Guangzhou,and the samples carrying ABCG2^(*)376T alleles were sequenced to confirm the accuracy of the HRM method.Results A HRM method,which can detect ABCG2^(*)376T allele and accurately type homozygotes and heterozygotes at the same time,had been established successfully.Fifteen individuals with heterozygous alleles were screened out of 1 560 blood donors in Guangzhou,while none homozygous allele was detected.Conclusion The HRM method can be used to accurately screen and type ABCG2^(*)376T allele.The frequency of this allele in Chinese population is about 0.48%(15/3120).

关 键 词：Jr(a-)血型 ABCG2^(*)376T等位基因 高分辨熔解曲线 分布频率 

分 类 号：R457.11[医药卫生—治疗学]