重组酶介导扩增技术与传统聚合酶链反应技术在甲状腺癌DNA甲基化检测中的应用比较  

Comparison of recombinase-aid amplification and traditional polymerase chain reaction in DNA methylation detection of thyroid cancer

作　　者：廖萍[1] 刘茶珍[1] 朱佩云[1] 刘国星[1] 王文静[1] 

机构地区：[1]上海市疾病预防控制中心公共卫生分子生物学研究室,200336

出　　处：《中国医药》2013年第6期797-799,共3页China Medicine

基　　金：上海市卫生系统优秀学科带头人培养计划资助（XBR201105I）

摘　　要：目的建立重组酶介导的核酸扩增（RAA）技术特异性检测DNA甲基化的新方法并与传统的DNA甲基化特异性PCR（MSP）方法进行比较。方法选取OXTR基因作为目的基因，提取样品外周血基因组DNA，经亚硫酸氢盐修饰后分别以MSP和RAA技术进行特异性检测DNA甲基化实验。结果2种技术皆能扩增出OXTR非甲基化条带，而RAA技术成功扩增OXTR甲基化条带。结论RAA是一种新型的等温体外核酸扩增技术，实现了在37℃恒温下的核酸快速扩增，可成为替代MSP乃至其他PCR实验的新方法。Objective To find a new technology and compare it with methylation specific polymerase chain reaction （MSP） in DNA methylation detection of thyroid cancer. Methods OXTR was selected as object of study. After samples DNA were extracted and were modified, the recombinase-aid amplification（RAA） and traditional MSP were separately used to amplify the target gene OXTR which was modified with bisulfite. Results Both the two technology succeeded in amplifying unmethylated OXTR gene. RAA succeeded in amplifying the methylated gene band. Conclusions RAA is a novel isothermal amplification technology in nucleic acid amplification technologies. It could be performed at 37 ℃ with no need to initial heat denaturation at a high temperature followed by amplification at a lower temperature, and this isothermal amplification technology may successfully compete with its widely used non-isothermal predecessor （PCR） for molecular biological study and application.

关 键 词：重组酶介导扩增技术 甲状腺癌 聚合酶链反应 DNA甲基化 

分 类 号：R736.1[医药卫生—肿瘤]