花生矮化病毒3种PCR检测方法的建立和比较

Development and Comparison of Three PCR Methods for Detecting Peanut stunt virus

建立了花生矮化病毒(PSV)普通RT-PCR、SYBR Green I荧光RT-PCR和免疫捕获RT-PCR检测方法,并比较了它们的检测灵敏度。结果表明,以阳性对照为样品,普通RT-PCR灵敏度达到10-4,SYBR Green I荧光RT-PCR和免疫捕获RT-PCR的检测灵敏度相当,相对灵敏度均可达到10-6;检测混有PSV的大豆叶片样品,普通RT-PCR灵敏度为10-1,SYBR Green I荧光RT-PCR和免疫捕获RT-PCR的检测灵敏度为10-5;SYBR Green I荧光RT-PCR和免疫捕获RT-PCR方法比直接RT-PCR灵敏度高至少100倍,尤其在带毒植物叶片检测中优势更明显,而且灵敏、简便、同时重复性好。

RT-PCR,SYBR Green I real-time RT-PCR and immune-capture RT-PCR approaches were developed for detection of Peanut stunt virus（PSV）,and their detecting sensitivity was compared.The results showed that the sensitivity of RT-PCR for detection of PSV was 10-4,whereas the sensitivity of SYBR Green I real-time RT-PCR and immune-capture RT-PCR was 10-6,respectively.The detection limit of RT-PCR for soybean leaf samples contaminated with PSV was 10-1,while that of SYBR Green I realtime RT-PCR and immune-capture RT-PCR for the same samples was 10-5,respectively.The detection sensitivity for PSV-contaminated samples by both SYBR Green I real-time RT-PCR and immune-capture RT-PCR was more than 100 times higher than that by RT-PCR,showing the favorable advantages of simplicity,high sensitivity and repeatability.