实时荧光定量PCR检测转基因玉米59122的方法建立及测量不确定度评估.pdfVIP

※分析检测 2014. 协，1.3工 NO.24 259 自由置迫害 实时荧光定量PCR检测转基因五米59122的 方法建主及测量不确定度评估 宋君，郭灵安气雷绍荣，王东，尹全，张富丽，刘文娟，常丽娟 (四川省农业科学院农产品质量标准研究所，四川成都 610066) 摘 要z 为了准确定量检测非故意扩散的转基因玉米59122，建立转基因玉米59122及产品的实时荧光定量聚合酶链 式反应 (polymerase chain reaction , PCR) 检测方法，并用特异性、准确度、灵敏度以及测量不确定度等指标评价 该方法。结果显示，利用该方法只能检出转基因玉米59122; 16 次重复测量含量为1%的转基因玉米59122样品，平均 测量值 (0.9%) 接近真实值(1%) ，相对误差为10%，测试回收率为师%，测量不确定度为0.002; 能检测到最低含量为 2 拷贝的59122分子片段:除去3 次偏离平均值较大的测量值外， 13 次重复测量值的变异系数为0.05。因此，实验建立的 转基因玉米59122实时荧光定量K双方法具有较高的特异性、准确度、灵敏度、精密度，较低的测量不确定度。 关键词:转基因玉米59122; 定量检测方法:方法学评价 An Accurate Quantitative Assay for Genetically Modified Maize Event 59122: Method Establishment and Assessment of Its Measurement Unce此ainty SONG Jun , GUO Ling-an *, LEI Shao-rong , WANG Dong, YIN Quan, ZHANG Fu心， LIU Wen-juan , CHANG Li才uan (lnstitute of Quality Standards and Testing Technology for Agro-Products , Sichuan Academy of Agricultural Sciences, Chengdu 61创始6， China) Abstract: Genetically modified maize (Event 59122) used as a feedstock has been approved to be imported to China since 2012. But, there has so far been no quantitative assay available to detect the genetically modified corn 59122 and its derivatives. In this study we established a quantitative detection method for this modified maize corn using real-time fluorescent q皿ntitative PCR in order to accurately examine whether it is spread unintentionally, ens田e 由e safety of corn production in China and reduce the ecological risk. Meanwhile, the method was evaluated by several methodological indicators such as specificity, sensitivity, accuracy and measurement uncertainty.