香蕉分化芽BBTV DAS-ELISA检测方法的建立和应用

摘要

为了解海南海口香蕉分化芽携带香蕉束顶病毒（Banana bunchy top virus ，BBTV）的情况，本研究建立了双抗夹心酶联免疫反应（DAS-ELISA）检测法，对来自海南地区组培厂的香蕉分化芽进行检测。检测结果表明，6个品种1852个香蕉分化芽平均带毒率为2．1％，其中‘皇帝蕉’为1．98％，‘粤科1号’为2．07％，‘广粉蕉’为1．98％，‘大蕉’为6．25％，‘218’为1．89％，‘巴西蕉’为2．25％。为了进一步验证 ELISA 结果的可靠性，随机选取12个阳性样品提取总 DNA 进行 PCR 鉴定。鉴定结果显示，12个样品中11个检测出有 BBTV，表明 DAS-ELISA 方法的准确率较高，与分子检测结果基本一致，可以胜任日常香蕉组培苗 BBTV 的检测。%In order to understand the situation of Banana bunchy top virus (BBTV)infection in banana differentiated shoots in Hainan,1 852 shoot samples from 6 banana varieties were detected by DAS-ELISAmethod.The results showed that the average BBTV-carrying rate was 2.1%,while 1.98% of ‘Huangdi’banana,2.07% of ‘Yueke No.1’banana, 1.98% of ‘Guangfen’banana,6.25% of Musa sapientum ,1.89% of ‘218’banana,and 2.25% of ‘Brazil’banana, respectively.To further confirm the accuracy of DAS-ELISA result,12 BBTV-carrying samples were selected for molec-ular detection by PCR.The results showed that 11 of them were BBTV positive,indicating DAS-ELISA method with the high accuracy can be used for BBTV detection in banana differentiated shoots.