用地高辛标记禽呼肠孤病毒(ARV)S1基因中编码σC蛋白的基因片段作为核酸探针,在斑点分子杂交中可检测到1.6pgARV的RNA。利用该核酸探针,通过检测鸡羽毛囊及体内病毒繁殖情况,比较研究了ARV感染后在鸡体内及鸡群中的传播动态。结果显示,研究建立的核酸探针检测方法灵敏度高、特异性强和操作简便,适于批量样品的检测。同时,用此方法检测发现,ARV感染24h后可侵染大部分器官,并且很快传播到同群未攻毒的鸡中,羽毛囊中的病毒检出率与鸡内脏器官中病毒的检出率一致。用核酸探针检测鸡羽毛囊中ARV的方法检测ARV的感染与流行情况,成本低,不影响鸡群生产。
The Avian reovirus (ARV) σC gene DNA fragment was labeled with Digoxigenin as cDNA probe for detection of ARV in chicken tissue and feather tips in Dot blot hybridization.And as little as 1.6 pg of ARV RNA could be detected with the DIG-labelled probe. ARV could be detected in most tissues of inoculated chickens 24 h post infection (PI) and spreaded to un-inoculated birds by contact rapidly. Positive rates in feather tips were correlated with the other tissues. The result demonstrated that the Dot blot with DIG probe to detect ARV in feather tips is a useful method for determination of ARV infection and spread in chickens, and it can be helpful to test a large number of samples.