禽传染性喉气管炎病毒TK基因狄高辛探针的研制及应用

Preparation and Application of Avian Infectious Laryngotracheitis Virus Thymidine Kinase Gene Digoxigenin labelled Probe

应用狄高辛标记禽传染性喉气管炎病毒（ＡＩＬＴＶ）ＴＫ基因制备探针，经斑点杂交显色后，探针同以ＡＩＬＴＶ北京株为模板的ＴＫ基因ＰＣＲ产物及北京株核酸均呈阳性紫色斑点，而与正常尿囊液、新城疫病毒和火鸡疱疹病毒、禽传染性支气管炎病毒无反应，２株确诊为ＡＩＬＴＶ的野外分离毒也呈阳性。本研究结果表明，该ＴＫ基因探针是敏感和特异的，可用于禽传染性喉气管炎和其它呼吸道疾病的鉴别诊断。

In this study, an avian infectious laryngotracheitis virus (AILTV) thymidine kinase gene (TK) probe labelled with digoxigenin was prepared, using dot blot hybridization, the probe showed strongly positive results when hybridized with a PCR product based on AILTV Beijing E2 strain and its template nucleic acid, but no hybridization signal was exhibited with other specimens such as normal allantoic fluid, NDV, HVT and AIBV. Two field samples identified to be AILTV wild stains gave result which corresponded to the positive hybridization reactions. The digoxigenin labelled AILTV TK gene probe was proved to be a highly sensitive and specific method of detection.