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PCR方法制备地高辛标记DNA探针检测中国对虾非包涵体型杆状病毒

徐洪涛;朴春爱;杨朵;王雷;张新宇;侯云德

  A pair of primers created from information of PmNOBⅢ genome DNA Sal I fragment produced a 355bp band by using Penaeus chinensis nonoccluded baculovirus (PcNOBV),the WSBV isolate from P.hinensis in mainland China,as the DNA template.The specific PCR product was cloned,sequenced and labeled with digoxigenin (DIG)DNA labeling kit(Boehringer Mannheim).The DIGlabeled fragment was tested by dotblot hybridization for sensitivity and specificity with purified PcNOBV nucleocapsid,PcNOBVinfected shrimp tissues and healthy shrimp tissues.The detection limit of the DNA probe is 6.8pg of purified PcNOBV DNA.No hybridization signals were observed using DNA from healthy shrimp as template.Healthy P.chinensis,artificially infected P.chinensis and pondreared adult P.chinensis were screened for PcNOBV infection by both PCR and the hybridization assay.The results showed a good relationship between PCR and the hybridization assay.These findings demonstrate that the DIGlabeled probe can be used as a sensitive,specific and costeffective reagent for detection of PcNOBV.……   
[关键词]:中国对虾;中国对虾非包涵体型杆状病毒(Pc NOBV);DNA探针;斑点杂交;聚合酶链反应(PCR)
[文献类型]:期刊
[文献出处]: 《病毒学报2000年01期
[格式]:PDF原版; EPUB自适应版(需下载客户端)