CAJ | 학술논문

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.
목적 건립중조매개도적핵산확증(RAA)기술특이성검측DNA갑기화적신방법병여전통적DNA갑기화특이성PCR(MSP)방법진행비교.방법 선취OXTR기인작위목적기인,제취양품외주혈기인조DNA,경아류산경염수식후분별이MSP화RAA기술진행특이성검측DNA갑기화실험.결과 2충기술개능확증출OXTR비갑기화조대,이RAA기술성공확증OXTR갑기화조대.결론 RAA시일충신형적등온체외핵산확증기술,실현료재37℃항온하적핵산쾌속확증,가성위체대MSP내지기타PCR실험적신방법.
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.