香蕉束顶病毒简易制样技术及分子检测

A Rapid Simple Sample Preparation and Molecular Detection of Banana Bunchy Top Virus

筛选了5对依据香蕉束顶病毒(BananaBunchyTopVirus,BBTV)外壳蛋白基因(CP)、复制酶基因(RF)保守区段设计的引物。结果显示,以RF为靶序列的引物,香蕉束顶病毒PCR特异性高,可以从感病组织中检测到ng(10-9g)级的病毒。样品提取缓冲液经过Sephadex凝胶介质过柱可有效浓缩病毒、去除PCR反应抑制物质,提高检测灵敏度,降低检测的假阴性。

Five primers designed according to conservative regions of coat protein gene （CP） and replicase gene （RP） of banana bunchy top virus were screened. The PCR amplification indicated that the primer RF1/RF2r based on RP sequence was specific and 10gg BBTV-infected tissue could be detected successfully. Additionally, the Sephadex gel filtration of extracted sap effectively concentrated the virus and eliminated the inhibitory substances of PCR reaction, and improved the detection sensitivity and decreased false negative of detection.