GICA-RT-PCR检测菜豆荚斑驳病毒的新方法  

A New Method for Detecting Bean Pod Mottle Virus by GICA-RT-PCR

作　　者：张晓雷[1] 檀根甲[1] 魏梅生[2] 张永江[2] 李桂芬[2] 

机构地区：[1]安徽农业大学植物保护学院,安徽合肥230063 [2]中国检验检疫科学研究院动植物检疫研究所,北京100029

出　　处：《大豆科学》2008年第6期1019-1023,共5页Soybean Science

基　　金：国家质量监督检验检疫总局资助项目(2006IK216)

摘　　要：菜豆荚斑驳病毒(Bean pod mottle virus,BPMV)是我国进境植物检疫性有害生物。为解决普通RT-PCR不能直接检测大豆病种子中该病毒的问题,将简便快速的胶体金免疫层析技术(GICA)和高灵敏度的普通RT-PCR检测技术有机地结合起来,建立了GICA-RT-PCR检测新方法。即先用GICA捕获病毒,将捕获的病毒直接进行RT-PCR扩增,简化了检测的步骤,提高了检测的灵敏度。结果表明:用该方法检测提纯病毒灵敏度达到0.01μg.mL-1、检测大豆病叶和病大豆种皮,灵敏度达到10-4,分别比GICA检测提高了20倍、10倍和100倍。GICA-RT-PCR可进一步确认GICA的结果。Bean pod mottle virus（ BPMV）is a quarantine pest for China. A high sensitive and rapid gold immunochromatography assay-RT-PCR（ GICA-RT-PCR ）method has been developed for the detection of BPMV- infected soybean seed. This method takes the advantage of gold immunochromatography assay （GICA）which captures the virus first on test line, then the immunocaptured virus will be used for direct RT- PCR to amplify the target nucleic acid, eliminating the need for traditional total RNA extraction methods and reducing the number of steps involved. The sensitivity of the GICA- RT- PCR for purified virus is 0.01 μg · mL^-1. BPMV- infected soybean leaves and soybean seed coat diluted to 10^-4 can be detected by GICA- RT-PCR. GICA-RT-PCR is 20 times, 10 times and 100 times more sensitive than GICA for detecting purified BPMV, BPMV- infected soybean leaves and soybean seed coat, respectively. GICA-RT-PCR is a simple and sensitive tool for confirmation of GICA results.

关 键 词：胶体金免疫层析 试纸条 菜豆荚斑驳病毒 GICA-RT-PCR 

分 类 号：S435.651[农业科学—农业昆虫与害虫防治] S41[农业科学—植物保护]