中文摘要：数字PCR技术作为转基因生物检测中的一项新技术，可测定靶标的绝对拷贝数。本文的研究目的是测试定量PCR（qPCR）技术与数字PCR（dPCR）技术的通用性，并通过开展两个不同dPCR平台性能的实验室分析，以测定参考物质中转基因含量的绝对GM拷贝数和GM拷贝数比。研究发现，两个dPCR系统中检测到的转基因比例均在可接受的变化范围内。然而，通过单个基因的绝对拷贝数可以发现其有三分之一表现出较高的变异性，这有可能是由于技术工作的变异性、液滴大小的变异性以及原始数据的分解而造成的。研究结果表明，qPCR与dPCR技术用于转基因定量检测是可比的。
外文摘要：Digital PCR (dPCR), as a new technology in the field of genetically modified (GM) organism (GMO) testing, enables determination of absolute target copy numbers. The purpose of our study was to test the transferability of methods designed for quantitative PCR (qPCR) to dPCR and to carry out an inter-laboratory comparison of the performance of two different dPCR platforms when determining the absolute GM copy numbers and GM copy number ratio in reference materials certified for GM content in mass fraction. Overall results in terms of measured GM% were within acceptable variation limits for both tested dPCR systems. However, the determined absolute copy numbers for individual genes or events showed higher variability between laboratories in one third of the cases, most possibly due to variability in the technical work, droplet size variability, and analysis of the raw data. GMO quantification with dPCR and qPCR was comparable. As methods originally designed for qPCR performed well in dPCR systems, already validated qPCR assays can most generally be used for dPCR technology with the purpose of GMO detection.
外文关键词：Digital PCR; Droplet digital PCR; Absolute quantification; Reference materials; GMO quantification
作者：Dobnik, David; Demsar, Tina; Huber, Ingrid; 等
作者单位：Natl Inst Biol NIB
期刊名称：ANALYTICAL AND BIOANALYTICAL CHEMISTRY
期刊影响因子：3.431
出版年份：2018
出版刊次：1
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