地高辛标记DNA探针对茶园NPV的检测

Detecting Nuclear Polyhedrosis Viruses in Tea Garden Using Digoxigenin Labeled DNA Probes

研究合成了茶尺蠖核型多角体病毒(Ectropis oblique nuclear polyhedrosis virus,EcobNPV)的地高辛标记DNA探针,采用斑点杂交检测茶园几种核型多角体病毒。结果表明,以含EcobNPV克隆片段的大肠杆菌菌液为样品时,斑点杂交检测灵敏度为80 CFU/μL。以感染病毒致死的茶尺蠖幼虫为试材,提取病毒基因组DNA,采用斑点杂交法检测均得到强的杂交信号。斑点杂交检测可有效区分茶园中几种核型多角体病毒,表明地高辛标记DNA探针的特异性好。

Digoxigenin-labeled DNA probes specific to Ectropis oblique nuclear polyhedrosis virus（EcobNPV） was synthetised,and several nuclear polyhedrosis viruses in tea garden were detected by dot blot hybridization.The results showed that the detection sensitivities of dot blot hybridization for Escherichia coli containing cloned fragments was 80 CFU/μL for EcobNPV.The dot blot hybridization method could specifically detect EcobNPV of larvae and give strong hybridization signal by extracting the DNA of viruses.The dot blot hybridization method could differentiate the viruses in tea garden,revealing that digoxigenin-labeled DNA probes has a high specificity.