一种5 重real-time PCR筛查转基因水稻方法的建立

以转基因水稻中最常用的CaMV35S启动子、NOS终止子、Cry1Ab/Ac基因、HPT基因及SPS水稻内标基因为研究对象，利用5 种不同的荧光信号（FAM、HEX、Taxas Red、Cy5、Cy5.5）进行多重实时聚合酶链式反应（real-time polymerase chain reaction，real-time PCR）检测方法的研究。通过引物组合筛选、反应体系优化、特异性测试、灵敏度测试、适用性测试等一系列实验，建立了5 重real-time PCR方法，灵敏度可达0.032%。此方法具有灵敏度高、结果准确、通量大等优点，可实现水稻中转基因成分的快速、高效检测。

Establishment of a Pentaplex Real-time PCR for Screening of Genetically Modified Rice

In this study, based on the CaMV35S promoter, the NOS terminator, Cry1Ab/Ac, HPT and the rice endogenous gene SPS, five different fluorescent probes (FAM, HEX, Taxas Red, Cy5, and Cy5.5) were used to develop and optimize a pentaplex real-time polymerase chain reaction (real-time PCR) method to detect genetically modified rice. For this purpose, a series of experiments was carried out to screen primer combinations, optimize the reaction system, and evaluate the specificity, sensitivity and applicability. The sensitivity of the proposed method was 0.032%. This method has the characteristics of high sensitivity, accuracy and high throughput and allows the rapid and efficient detection of genetically modified components in rice.