中文摘要：商业上可获得的转基因植物在世界范围内被认可，因此转基因生物分子检测的目标序列不断增加。欧盟已经实施了一套非常严格的程序以批准种植、进口和/或利用转基因生物作为食品或食品成分。因此，GMO实验室和食品生产行业目前被迫采用不同的方法来测试原材料和复杂的加工食品。三个外源基因（35 S启动子、NOS终止子和新霉素磷酸转移酶II）通常用于转基因检测。本文建立了一个多重定量实时PCR（QPCR）系统，该系统能够在一个反应管中同时检测三个外源基因。在玉米样品中多重qPCR检测的确定限为4 copies/reaction。根据23个转基因作物（GM）和97个复合加工食品的检测结果发现，该方法检测的特异性为100%。研究结果表明，与参考基因结合的高通量多重QPCR方法对于筛选转基因生物甚至加工食品都是可行的。
外文摘要：The commercially available genetically modified plants authorized worldwide and therefore the target sequences for molecular detection of genetically modified organisms (GMOs) are ever-increasing. The European Union has implemented a set of very strict procedures for approval to grow, import and/or utilize GMOs as food or food ingredients. As a result, GMO laboratories and food production industry currently are forced to apply different methods to test raw material and complex processed food products. Three exogenous genes (the 35 s promoter of the cauliflower mosaic virus (35 s), nos terminator from Agrobacterium tumefaciens (nos), and the neomycin phosphotransferase II (nptII) gene) are commonly used in GMO detection. In this paper, a multiplex quantitative real-time PCR (qPCR) system was developed which allows simultaneously detection of the three exogenous genes in one reaction tube. The determined limits for the multiplex qPCR assays were 4 copies/reaction in maize samples. The specificity of the assays was demonstrated to be 100% according to the detection results of 23 genetically modified (GM) crops and 97 complex processed food products. The validation data show the individual PCR efficiency was accredited with negligible impacts between three detection channels in 7500 fluorescence quantitative PCR machine. These results indicate that this high-throughput multiplex qPCR method which combined with a reference gene is feasible for screening of GMOs, even for the processed food.
外文关键词：Detection, Genetically modified organisms, Multiplex qPCR, Screening
作者：Wang, Fengjun; Feng, Junli; Ye, Sudan; Huang, Hannian; Zhang, Xianglin
作者单位：Zhejiang Gongshang Univ
期刊名称：BIOLOGIA
期刊影响因子：2.0
出版年份：2018
出版刊次：1
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