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基于数字PCR的单分子DNA定量技术研究进展
引用本文:李亮,隋志伟,王晶,臧超,余笑波.基于数字PCR的单分子DNA定量技术研究进展[J].生物化学与生物物理进展,2012,39(10):1017-1023.
作者姓名:李亮  隋志伟  王晶  臧超  余笑波
作者单位:中国计量科学研究院;浙江省计量科学研究院
基金项目:国家科技支撑计划资助项目(2008BAK41B01)
摘    要:数字PCR是一项针对单分子目标DNA的绝对定量技术.该技术是将含有DNA模板的反应溶液分配到大量独立的反应室中并且发生扩增反应,通过统计反应室中的阳性信号来定量DNA的拷贝数.DNA样品在反应室中随机和独立分布是单分子成功扩增和准确定量DNA拷贝数的关键因素.本文综述了数字PCR的发展历史、数字PCR与实时荧光定量PCR的区别,以及数字PCR在临床诊断、转基因成分定量、单细胞基因表达、环境微生物检测和下一代测序等方面的最新进展,并展望了该技术的应用前景.

关 键 词:数字PCR,绝对定量,实时荧光定量PCR,拷贝数,单分子
收稿时间:1/3/2012 12:00:00 AM
修稿时间:3/8/2012 12:00:00 AM

Progress of Digital PCR for Single DNA Quantification
LI Liang,SUI Zhi-Wei,WANG Jing,ZANG Chao and YU Xiao-Bo.Progress of Digital PCR for Single DNA Quantification[J].Progress In Biochemistry and Biophysics,2012,39(10):1017-1023.
Authors:LI Liang  SUI Zhi-Wei  WANG Jing  ZANG Chao and YU Xiao-Bo
Institution:1) National Institute of Metrology,Beijing 100013,China;2) Zhejiang Institute of Metrology,Hangzhou 310013,China)
Abstract:Digital PCR is an absolute DNA quantification technique to determine the copy number of target DNA. PCR reaction solution is divided into aliquot throughout numerous partitions and amplifies independently. The copy number of target DNA is estimated by the statistical analysis of positive signals. Random and independent distribution of the target DNA molecules throughout the partitions of the digital panel and successful amplification from single molecules are critical to the validity of this approach for estimating target DNA copy number. In this review, we discuss the advances in development of digital PCR, the differences between digital PCR and real time quantitative PCR. We further show the advances in application of digital PCR in the clinical diagnosis, GMO analysis, single cell expression, environmental microbiology and next generation sequencing. The prospects for the applications of digital PCR are discussed.
Keywords:digital PCR  absolute quantification  real time quantitative PCR  copy number  single molecular
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