低温变性下复合PCR技术及其应用  

Co-amplification at lower denaturation temperature-PCR:methodology and applications

作　　者：梁卉[1,2] 陈国杰 于燕 熊礼宽[1,2] Hui Liang;Guojie Chen;Yan Yu;Likuan Xiong(Central Laboratory, Bao’an Maternal and Child Health Hospital, Jinan University, Shenzhen 518102, China;Shenzhen Key Laboratory of Birth Defects, Shenzhen 518102, China;Department of Gastroenterology, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China;Department of Obstetrics, Bao’an Maternal and Child Health Hospital, Jinan University, Shenzhen 518102, China)

机构地区：[1]暨南大学附属深圳市宝安区妇幼保健院中心实验室,深圳518102 [2]深圳市出生缺陷重点实验室,深圳518102 [3]郑州大学附属第一医院消化科,郑州450052 [4]暨南大学附属深圳市宝安区妇幼保健院产科,深圳518102

出　　处：《遗传》2018年第3期227-236,共10页Hereditas（Beijing)

基　　金：深圳市三名工程-出生缺陷防治研究与转化团队(编号:SZSM201406007);深圳市出生缺陷重点实验室(编号:ZDSYS201504301707152);深圳市宝安区医疗卫生基础研究项目(编号:2014067,2017JD001)资助

摘　　要：低温变性下复合PCR(co-amplification at lower denaturation temperature-polymerase chain reaction,COLDPCR)是一种在高丰度野生型序列背景下选择性变性和扩增低丰度突变型序列的方法,可将突变型序列富集10~100倍。基于突变片段Tm值的改变和异源双链DNA分子的形成,COLD-PCR可富集扩增片段中所有类型和位置的突变,也可富集未知突变,具有敏感、特异、精确、廉价和易操作等优点。COLD-PCR及其衍生方法被应用于肿瘤、微生物、产前筛查和动植物等领域,对疾病的早期诊断、病程和治疗监控、药物选择、预后判断和植物育种等均有积极的作用。本文就COLD-PCR的原理、关键技术、衍生方法及其应用进行综述。Co-amplification at lower denaturation temperature-polymerase chain reaction(COLD-PCR)is a novel form of PCR that selectively denatures and amplifies low-abundance mutations from mixtures of wild-type and mutation-containing sequences,enriching the mutation10to100folds.Due to the slightly altered melting temperature(Tm)of the dou ble-stranded DNA and the formation of the mutation/wild-type heteroduplex DNA,COLD-PCR methods are sensitive,specific,accurate,cost-effective and easy to maneuver,and can enrich mutations of any type and at any position,even unknown mutations within amplicons.COLD-PCR and its improved methods are now applied in cancer,microorganisms,prenatal screening,animals and plants.They are extremely useful for early diagnosis,monitoring the prognosis of disease and the efficiency of the treatment,drug selection,prediction of prognosis,plant breeding and etc.In this review,we introduce the principles,key techniques,derived methods and applications of COLD-PCR.

关 键 词：低温变性下复合PCR 精确的变性温度 选择性富集 低丰度突变 检测阈值 

分 类 号：Q503[生物学—生物化学]