转基因棉花MON88701品系特异性实时荧光PCR检测方法的建立

【目的】建立转基因棉花MON88701品系特异性实时荧光聚合酶链反应(polymerase chain reaction,PCR)检测方法。【方法】利用实时荧光PCR技术,根据转基因棉花MON88701品系特异性序列设计引物和探针,建立转基因棉花MON88701实时荧光PCR检测方法,并测定本方法的灵敏度、特异性及可重复性。【结果】建立的检测方法特异于转基因棉花MON88701成分的检测,灵敏度测试表明其定量下限为34拷贝;重复性试验显示其相对标准偏差在可接受范围内。【结论】本研究建立的转基因棉花MON88701品系特异性实时荧光PCR检测方法具有良好的特异性和高灵敏度,适合对转基因棉花MON88701品系进行检测。

Establishment event-specific real-time polymerase chain reaction method for detection of genetically modified cotton line MON88701

Aim] To establish a real-time PCR detection method for the genetically modified (GM) Roundup Ready cotton event MON88701, which has not been authorized by the Ministry of Agriculture of China.Method] The specific primer pairs and probe based on the MON88701 event-specific sequence were designed and then the real-time PCR detection system was established. The specificity, sensitivity and repeatability were analyzed.Result] The real-time PCR method was specific for detecting MON88701. The limit of quantification (LOQ) was 34 copies MON88701 genomic DNA. Repeatability of the established event-specific real-time PCR method was assessed and the relative standard deviation (RSD) was within the acceptable range.Conclusion] The established event-specific quantitative real-time PCR method of MON88701 in this study has a good specificity and high sensitivity, and is suitable for the detection of the unathorised event MON88701.