遗传 ›› 2020, Vol. 42 ›› Issue (4): 363-373.doi: 10.16288/j.yczz.19-351

• 综述 • 上一篇    下一篇

数字PCR技术及其在检测领域的应用

冯秀晶, 易红梅, 任星旭, 任佳丽, 葛建镕, 王凤格()   

  1. 北京市农林科学院玉米研究中心,玉米DNA指纹及分子育种北京市重点实验室,北京 100097
  • 收稿日期:2019-11-18 修回日期:2020-03-12 出版日期:2020-04-20 发布日期:2020-03-31
  • 通讯作者: 王凤格 E-mail:gege0106@163.com
  • 作者简介:冯秀晶,博士,研究方向:转基因植物检测。E-mail: fengxj187@126.com;|易红梅,博士,副研究员,研究方向:玉米分子鉴定。E-mail: yhmmaize@163.com; 冯秀晶和易红梅为并列第一作者。
  • 基金资助:
    国家重点研发计划项目编号(2017YFD0102001);转基因生物新品种培育重大专项资助编号(2017ZX08013001)

Digital PCR and its application in biological detection

Feng Xiujing, Yi Hongmei, Ren Xingxu, Ren Jiali, Ge Jianrong, Wang Fengge()   

  1. Beijing Key Laboratory of Maize DNA Fingerprinting and Molecular Breeding, Maize Research Center, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100097, China
  • Received:2019-11-18 Revised:2020-03-12 Online:2020-04-20 Published:2020-03-31
  • Contact: Wang Fengge E-mail:gege0106@163.com
  • Supported by:
    Supported by the National Key Research and Development Program of China No(2017YFD0102001);the National Genetically Modified Organisms Key Breeding Projects of China No(2017ZX08013001)

摘要:

随着分子生物学技术的不断发展和需求的多样化,用于核酸检测的各种PCR衍生技术应运而生。数字PCR是一种单分子水平的大规模分区扩增定量核酸检测技术。该技术以微腔室/微孔或微滴作为PCR反应器,无需校准物和绘制标准曲线即可实现对样品初始浓度的绝对定量,具有高灵敏度、高特异性和高精确度的特点。本文详细介绍了数字PCR的技术发展史、作用原理以及仪器平台类型,系统阐述了数字PCR在转基因检测、疾病诊断、环境及食品监管等方面的应用概况,并对该技术的应用前景进行了展望,以期对未来数字PCR的开发利用提供参考。

关键词: 数字PCR, 单分子, 绝对定量

Abstract:

Various derivative technologies based on PCR for nucleic acid detection have emerged with the continuous development and the diverse needs of molecular biology technology. Digital PCR (dPCR) is a nucleic acid detection method for large scale amplification based on a single molecular template, which runs an individual PCR reaction using chambers/wells or droplets. dPCR can be used for absolute quantification for the initial concentration of samples without calibrator and drawing standard curve, showing the characteristics of high sensitivity, specificity, and accuracy. In this review, we introduce the history of technology development, principle, and instrument platform types of digital PCR in detail. Then, we summarize the application of this technology in GMO quantification, disease diagnosis, environment and food supervision. Finally, we describe the application prospect of dPCR, providing a reference for the development and utilization of this technology in the future.

Key words: digital PCR, single molecule, absolute quantification