利用ELISA检测两种兰花病毒的研究  

Detection of two orchid viruses using ELISA

作　　者：柳爱春[1] 刘超[1] 赵芸[1] 徐晓丹[1] 

机构地区：[1]杭州市农业科学研究院,浙江杭州310024

出　　处：《浙江农业学报》2009年第2期91-95,共5页Acta Agriculturae Zhejiangensis

基　　金：杭州市科技发展计划项目(20070132N09)

摘　　要：建立了三抗夹心酶联免疫吸附法(TAS-ELISA)检测建兰花叶病毒(Cymbidium mosaic virus,CymMV)和双抗夹心酶联免疫吸附法(DAS-ELISA)检测齿兰环斑病毒(Odontoglossum ringspot virus,ORSV)的方法。共检测了浙江省内139个兰科样品和8个百合科样品,结果表明有31.7%的兰花检出CymMV,有28.8%的兰花检出ORSV。根据病毒外壳蛋白基因上下游保守序列设计了检测ORSV的引物,参照周国辉报道设计了检测CymMV的引物并对人工接种两种病毒的兰花进行了逆转录聚合酶链式反应(RT-PCR)检测,结果表明,RT-PCR检测结果与ELISA结果相一致,进一步证实本研究建立的TAS-ELISA,DAS-ELISA方法稳定可靠,重复性好,可应用于兰花上CymMV和ORSV两种病毒的早期诊断和检测。The methods of TAS-ELISA and DAS-ELISA were established to detect Cymbidium mosaic virus （CymMV） and Odontoglossum ringspot virus （ORSV）, respectively. The experiment was carried out with 139 orchidaceous samples and 8 liliaceous samples in Zhejiang province. The results showed that 31.7% of the orehidaceous samples were infected with CymMV, 28.8% of the orchidaceous samples were infected with ORSV.According to the conserved sequences of coat pro- tein-encoding genes （CP） of ORSV and the report of Zhou et al, the specific primers were designed to detect CymMV and ORSV by RT-PCR from the orchid infected with viruses artificially. The results of RT-PCR was consistent with those of ELISA, which suggested that the methods of TAS-ELISA, DAS-ELISA established here, were reliable and repeatable, and could be used for early diagnosis and detection of CymMV and ORSV in orchid.

关 键 词：建兰花叶病毒 齿兰环斑病毒 TAS-ELISA DAS-ELISA 一步法RT-PCR 

分 类 号：S682.31[农业科学—观赏园艺] S432.41[农业科学—园艺学]