实时荧光定量PCR方法检测转基因玉米MIR604

A real-time fluorescent quantitative PCR for detection of genetically modified maize line MIR604

转基因玉米MIR604是瑞士先正达公司2005年研发的抗虫玉米,中国于2008年批准MIR604进口作为食品和饲料的加工原料。为了精确定量检测MIR604及其加工产品,在MIR604的左侧边界序列和玉米基因组之间设计引物和探针,成功建立了MIR604品系特异实时荧光定量PCR检测方法。采用该方法设计的引物和探针,对6种非转基因作物、MIR604及其他非目标转基因作物进行检测,除MIR604外,其余样品均未检测到MIR604分子片段的扩增信号;对已知含量为1%的MIR604标准品进行定量检测,测量值为1.07%,接近真实值1%;灵敏度实验结果显示,该方法能检测到仅5个拷贝的MIR604分子片段。该试验所建立的MIR604品系特异实时荧光定量PCR检测方法具有较高的特异性、准确度及灵敏度,可用于转基因玉米MIR604的定量检测。

Genetically modified maize MIR604 is an insect resistant maize line developed by Syngenta in 2005,which was approved as a raw material for food and feed processing in 2008 in China. In order to accurately detect MIR604 and its processing products,event-specific quantitative PCR detection method was established in this study.The PCR primers and probes were designed specially according to the left boundary sequence of MIR604 and maize genome sequence. Finally,we used the method to detect 6 kinds of non-genetically modified crops,MIR604 and other non-target transgenic crops. The results showed that the other samples apart from MIR604 were not detected. The measured value of the sample（ CRM） containing 1% MIR604 was 1. 07%,which was close to the real value（ 1%）.The sensitivity experiment results indicated the method could detect only 5 copies of MIR604 molecular fragments.Thus,the event-specific quantitative PCR detection method of genetically modified maize MIR604 has high specificity,accuracy and sensitivity,which can be used for quantitative detection of genetically modified maize MIR604.