鸡致病性外源性禽白血病病毒特异性核酸探针交叉斑点杂交检测试剂盒的研制

Cross Dot Blot Hybridization with Specific Nucleic Acid Probesfor Detection of Pathogenic Exogenous Avian Leucosis Virus

关键词:  鸡致病性外源禽白血病病毒  鸡内源性禽白血病病毒  特异性核酸探针  PCR  交叉斑点分子杂交  试剂盒

Key words:  pathogenic exogenous ALV  endogenous ALV  specific nucleic acid probe  PCR  cross dot blot hybridization

摘要:

用特定引物通过PCR合成了经地高辛标记的鸡致病性外源性及内源性禽白血病病毒特异性核酸探针,通过交叉斑点分子杂交,这些探针将可用于检测病料样品中致病性外源性禽白血病毒特异性核酸的存在。利用此试剂盒,对从病料组织样品中提取的基因组DNA作交叉斑点分子杂交或对提取的DNA用相应引物扩增后的PCR产物作交叉斑点分子杂交,可在24~36 h内完成检测并报告结果。

Abstract:

Four DIG-labeled nucleic acid probes for specific detection of chicken endogenous and pathogenic exogenous avian leukosis virus(ALV) were synthesized by PCR using specific primers, these probes are used to detect exogenous pathogenic ALV specific nucleic acid in suspected samples through cross dot hybridization. Genomic DNA extracted from the samples or its amplified PCR products using the appropriate primers can be detected with specific probes. The results could be reported in 24 ~ 36h by cross-dot hybridization.