数字PCR技术在核酸标准物质研制中的应用  

Application of Digital PCR Technology in the Development of Nucleic Acid Reference Materials

作　　者：郑子繁 柳方方[1] 刘卫晓 金芜军[1] 李亮[1] ZHENG Zifan;LIU Fangfang;LIU Weixiao;JIN Wujun;LI Liang(Institute of Biotechnology,Chinese Academy of Agricultural Sciences,Beijing 100081,China)

机构地区：[1]中国农业科学院生物技术研究所,北京100081

出　　处：《生物技术进展》2020年第6期579-584,共6页Current Biotechnology

基　　金：国家转基因新品种培育重大专项(2016ZX08012-003);中央级公益性科研院所基本科研业务费专项(16103920200010)。

摘　　要：在标准物质研制领域,生物标准物质的研制逐渐成为了研究热点,同时基于核酸的检测技术的开发与应用又推动了核酸标准物质的进程。核酸标准物质需要高级别、精准的定值方法,数字PCR作为单分子定量技术得到了广泛的应用。数字PCR是一种测定核酸分子的绝对定量方法,如微滴式数字PCR是采用油包水形成的微滴作为反应室,将含有DNA模板的反应溶液分配到大量独立的反应室中进行扩增反应,再通过统计反应室中的阳性信号来定量DNA的拷贝数,从而达到精确定量核酸拷贝数的目的。综述了近年来关于数字PCR及其在核酸标准物质研究领域的最新应用进展,重点综述了其在转基因检测、医疗诊断等领域的应用进展,以期为核酸标准物质的研制提供参考。In the field of reference material development,the development of biological reference material has gradually become a hot topic.At the same time,the development and application of nucleic acid-based detection technology has promoted the process of nucleic acid reference material.Nucleic acid reference materials require high-level and accurate valuation methods,and digital PCR has been widely used as a single-molecule quantitative technology.Digital PCR is an absolute quantitative method for the determination of nucleic acid molecules.For example,droplet digital PCR uses droplets formed by water-in-oil as the reaction chamber,and distributes the reaction solution containing the DNA template to a large number of independent reaction chambers for expansion.Increase the reaction,and then quantify the copy number of DNA by counting the positive signals in the reaction chamber,so as to achieve the purpose of accurately quantifying the copy number of nucleic acid.This article reviewed the literature on the research of digital PCR and the latest application progress of digital PCR technology in the development of DNA and nucleic acid reference materials,especially in the fields of medical diagnosis and genetic modification detection,which was expected to provide reference for the research of reference material development.

关 键 词：数字PCR 标准物质 生物标准物质 测量不确定度 

分 类 号：Q52[生物学—生物化学]