中文摘要：用数字PCR可以实现对转基因生物品种的绝对定量检测，但大多数基于数字PCR的绝对定量方法需要预处理步骤，同时单一检测无法满足基于外源片段和内参基因比的转基因品种绝对定量的需求。因此，为了将数字PCR用于不同转基因生物品种的绝对定量检测，本文开发了一种无需预处理的、基于双联室数字PCR的定量检测方法。作者用该方法检测了7个转基因品种进行实证分析，并用定量限（LOQ）、检测限（LOD）和特异性指标进行验证。研究结果发现，与单一数字PCR相比，双联室数字PCR的RSD检测结果更低；双联室数字PCR检测系统是用一个简单而稳定的方式来实现不同转基因生物品种的绝对定量检测；此外，LOQ和LOD指标表明该方法适用于转基因品种的日常检测和定量分析。
外文摘要：The possibility of the absolute quantitation of GMO events by digital PCR was recently reported. However, most absolute quantitation methods based on the digital PCR required pretreatment steps. Meanwhile, singleplex detection could not meet the demand of the absolute quantitation of GMO events that is based on the ratio of foreign fragments and reference genes. Thus, to promote the absolute quantitative detection of different GMO events by digital PCR, we developed a quantitative detection method based on duplex digital PCR without pretreatment. Moreover, we tested 7 GMO events in our study to evaluate the fitness of our method. The optimized combination of foreign and reference primers, limit of quantitation (LOQ), limit of detection (LOD) and specificity were validated. The results showed that the LOQ of our method for different GMO events was 0.5%, while the LOD is 0.1%. Additionally, we found that duplex digital PCR could achieve the detection results with lower RSD compared with singleplex digital PCR. In summary, the duplex digital PCR detection system is a simple and stable way to achieve the absolute quantitation of different GMO events. Moreover, the LOQ and LOD indicated that this method is suitable for the daily detection and quantitation of GMO events. (C) 2016 Elsevier B.V. All rights reserved.
外文关键词：Absolute quantitation；Digital polymerase chain reaction；Duplex detection；Pretreatment-free
作者：Zhu, Pengyu；Fu, Wei；Wang, Chenguang；等
作者单位：中国农业大学
期刊名称：ANALYTICA CHIMICA ACTA
期刊影响因子：4.712
出版年份：2016
出版刊次：4
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