PCR结合斑点杂交技术在检测禽网状内皮增生病毒中的应用

为评价PCR结合斑点杂交技术在鸡REV检测中的应用价值,采用PCR法制备禽网状内皮细胞增生症病毒特异性地高辛标记DNA探针,同时用PCR技术、斑点杂交方法和PCR产物斑点杂交方法检测了不同地区的病、死鸡的组织样品REV的感染情况。结果表明,PCR产物斑点杂交法的检出率(45.16%,14/31)高于组织DNA直接斑点杂交法(32.26%,10/31),显著高于单纯PCR扩增法(0%,0/31)。PCR结合斑点杂交检测技术能快速、敏感、准确,充分避免PCR中的假阴性和假阳性现象,值得推广应用。

Use of PCR and Dot Blot Hybridization to Detect Chicken Reticuloendotheliosis Virus

In order to evaluate the combination of PCR and dot blot assay in detection for REV infection in chicken samples,the REV-specific digoxigenin-labled DNA probe was prepared by PCR.DNA samples were extracted and tested by regular PCR and dot blot hybridization respectively or by PCR plus dot blot.The sensitivity of different methods was compared with each other.Results showed that the positive rate（45.16%,14/31） using PCR-Dot blot was higher than that（32.26%,10/31） by direct dot blot for tissue DNA,and significantly higher than that（0%,0/31） by PCR amplification.The combined PCR-Dot blot assay is a rapid,sensitive and specific diagnosis method.It could overcome false negative and false positive in PCR,and the sensitivity was significantly improved.