禽弱毒疫苗中鸡传染性贫血病病毒nested-PCR检测方法的建立

发布时间：2019-04-27 15:04

【摘要】：疫苗中污染鸡传染性贫血病毒(CIAV)是造成CIAV感染的途径之一,而通常疫苗中CIAV污染剂量较低,应用一般的技术难以检测到。为了更灵敏地检测禽弱毒疫苗中CIAV的污染,根据Gen Bank已发表的CIAV基因序列,针对其保守区设计了外用引物和内用引物,通过优化反应体系和反应条件建立了检测CIAV的nested-PCR检测方法。结果显示,所建立的检测方法灵敏度比常规PCR高100倍,该方法可以检测到1 000羽份弱毒疫苗中1 EID_(50)的低剂量CIAV污染,应用该方法从送检的50个疫苗样品中检测出4个样品为CIAV阳性。该方法与禽白血病病毒、禽网状内皮组织增生症病毒以及马立克氏病病毒等的基因均无交叉反应,具有很好的特异性。提示所建立的nested-PCR灵敏度高、特异性强,可用于检测禽弱毒疫苗中CIAV低剂量的污染。
[Abstract]:The contamination of chicken infectious anemia virus (CIAV) in vaccines is one of the ways to cause CIAV infection. However, the contaminated doses of CIAV in vaccines are usually low, so it is difficult to detect them by common techniques. In order to detect the contamination of CIAV in attenuated avian vaccine more sensitively, according to the published CIAV gene sequence of Gen Bank, two primers were designed for the conservative region of the vaccine, external primer and internal primer. The nested-PCR detection method for CIAV was established by optimizing the reaction system and reaction conditions. The results showed that the sensitivity of the established method was 100 times higher than that of conventional PCR. The method could detect low dose CIAV contamination in 1 000 EID_ (50) attenuated vaccine. The method was used to detect CIAV positive in 4 samples of 50 vaccine samples. The method has no cross-reaction with the genes of avian leukemia virus, reticuloendotheliosis virus and Marek's disease virus, and has good specificity. It is suggested that the established nested-PCR has high sensitivity and specificity, and can be used to detect low-dose contamination of CIAV in avian attenuated vaccine.
【作者单位】：山东农业大学动物医学院;
【基金】：农业部财政专项“种禽场主要垂直传播性疾病疫情调查”项目
【分类号】：S858.31

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