中文摘要：市场上转基因生物的数量正在稳步增长。目前，基于DNA的检测方法比基于蛋白质的检测方法更受欢迎，而实时定量PCR（qPCR）仍然是转基因生物分析中的黄金标准。而数字PCR（DdPCR）比QPCR多了几个优点，使得这种新技术也适用于转基因生物分析。本文研究重点是研究使用DPCR进行转基因生物量化的目的和参数，例如DNA的质量和纯度、反应优化等，这对实现精确和可靠的结果是很重要的。本文对三个关键因素进行了更深入的探讨和讨论：正确划分分区为正，正确确定分区体积，稀释因子。本文研究可以作为所有实验室实施DPCR的指南，且大多数讨论的参数适用于除纯GMO测试以外的字段。
外文摘要：The number of genetically modified organisms (GMOs) on the market is steadily increasing. Because of regulation of cultivation and trade of GMOs in several countries, there is pressure for their accurate detection and quantification. Today, DNA-based approaches are more popular for this purpose than protein-based methods, and real-time quantitative PCR (qPCR) is still the gold standard in GMO analytics. However, digital PCR (dPCR) offers several advantages over qPCR, making this new technique appealing also for GMO analysis. This critical review focuses on the use of dPCR for the purpose of GMO quantification and addresses parameters which are important for achieving accurate and reliable results, such as the quality and purity of DNA and reaction optimization. Three critical factors are explored and discussed in more depth: correct classification of partitions as positive, correctly determined partition volume, and dilution factor. This review could serve as a guide for all laboratories implementing dPCR. Most of the parameters discussed are applicable to fields other than purely GMO testing.
外文关键词：Digital PCR, Droplet digital PCR, Chip-based digital PCR, Genetically modified organisms, Quantification
作者：Demeke, Tigst; Dobnik, David
作者单位：Natl Inst Biol
期刊名称：ANALYTICAL AND BIOANALYTICAL CHEMISTRY
期刊影响因子：3.307
出版年份：2018
出版刊次：7
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