转基因大豆MON89788实时荧光PCR检测方法的建立

Establishment of Event-Specific Real-Time PCR Method for Detection of Genetically Modified Soybean MON89788

为实现转基因大豆MON89788的标识管理,针对转基因大豆MON89788的品系特异性序列设计引物和Taq Man探针,建立转基因大豆MON89788实时荧光聚合酶链式反应(polymerase chain reaction,PCR)检测方法,并对该方法的特异性、灵敏度和重复性进行检测。结果显示:建立的转基因大豆MON89788实时荧光PCR检测方法能扩增出127 bp的产物,特异性强,灵敏度达到0.1%,约为40个单倍体基因组拷贝,检测重复性好,可成功应用于实际样品检测。因此,建立的转基因大豆MON89788实时荧光PCR检测方法可以应用于转基因大豆MON89788大豆及其制品的检测。

For implementation of labeling regulations, an event-specific real-time PCR method for the detection of genetically modified soybean MON89788 was established in this study. Primers and Taq Man probe were designed based on the event-specific sequence of MON89788 soybean. The specificity, sensitivity and repeatability of the developed method were examined, respectively. The results showed that the method could amplify a 127 bp product specifically; the limit of detection was 0.1% in 50 ng of soybean genomic DNA, approximately corresponding to 40 soybean haploid genome copies; and this method had good repeatability and could be successfully applied to the detection of real samples. These results indicated that the strain-specific real-time PCR method was suitable for the identification of genetically modified soybean MON89788 and its products.