Tm-shift基因分型方法在遗传学中的应用  

Application of Tm-shift genotyping method in genetic studies

作　　者：袁芳[1] 徐进[1,2] 季林丹[1,2] 费丽娟[1] 刘盼盼[1] 张莉娜[1] 

机构地区：[1]宁波大学医学院,宁波315211 [2]中国科学院昆明动物研究所-香港中文大学生物资源与疾病分子机理联合实验室,昆明650223

出　　处：《遗传》2012年第11期1484-1490,共7页Hereditas（Beijing)

基　　金：浙江省自然科学基金项目(编号:(Y2100857);浙江省教育厅重点科研项目(编号:Z201017918);宁波市自然科学基金项目(编号:2012A610237;2011A610037)资助

摘　　要：Melting Temperature shift(Tm-shift)是一种新的基因分型方法,主要通过在两条特异性引物5′端加入不同长度的GC序列,PCR扩增后根据熔解曲线中产物Tm值的差异来完成分型。文章建立了Tm-shift法对2 048份样品的29个SNP进行分型,通过分型成功率、重复检测一致率、测序验证准确度综合评价分型效果。结果显示,29个SNP中有27个可以采用本方法分型,分型成功率为93.1%。测序验证准确性达到100%。3种基因型阳性标准对照重复检测一致率为100%;100个随机样品重复检测,重复性为97%。因此,Tm-shift基因分型法是一种成本低廉、准确灵敏、稳定可靠、通量灵活、操作简便的基因分型方法,可在遗传学研究中推广应用。Melting Temperature shift （Tm-shift） is a new genotyping method. With two GC-rich tails of unequal length combined to 5＇-terminal of allele-specific primers, genotypes can be determined by the distinct Tms of the PCR products with inspection of a melting curve on the real-time PCR machine. In this study, 29 SNPs were genotyped with 2 048 sam- ples by using Tm-shift genotyping method, and the results were assessed by success rate, consistent rate, and accuracy. The results indicated that among 29 SNPs, 27 SNPs could be genotyped by Tm-shift. In other words, the success rate was 93.1% The accuracy confirmed by direct sequencing was 100%. The consistency was 100% with 3 control samples, and 97% from a replication study in 100 samples. Thus, Tm-shift is a genotyping method with advantages including low cost, high accu- racy, stability, reliability, flexible throughput, and easy operation, which can be applied to genetic studies widely.

关 键 词：Tm-shift 基因分型 遗传学 

分 类 号：R450[医药卫生—治疗学]