【摘要】：疫苗中污染雞傳染性貧血病毒(CIAV)是造成CIAV感染的途徑之一,而通常疫苗中CIAV污染劑量較低,應(yīng)用一般的技術(shù)難以檢測到。為了更靈敏地檢測禽弱毒疫苗中CIAV的污染,根據(jù)Gen Bank已發(fā)表的CIAV基因序列,針對其保守區(qū)設(shè)計了外用引物和內(nèi)用引物,通過優(yōu)化反應(yīng)體系和反應(yīng)條件建立了檢測CIAV的nested-PCR檢測方法。結(jié)果顯示,所建立的檢測方法靈敏度比常規(guī)PCR高100倍,該方法可以檢測到1 000羽份弱毒疫苗中1 EID_(50)的低劑量CIAV污染,應(yīng)用該方法從送檢的50個疫苗樣品中檢測出4個樣品為CIAV陽性。該方法與禽白血病病毒、禽網(wǎng)狀內(nèi)皮組織增生癥病毒以及馬立克氏病病毒等的基因均無交叉反應(yīng),具有很好的特異性。提示所建立的nested-PCR靈敏度高、特異性強,可用于檢測禽弱毒疫苗中CIAV低劑量的污染。
[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.
【作者單位】： 山東農(nóng)業(yè)大學(xué)動物醫(yī)學(xué)院;
【基金】：農(nóng)業(yè)部財政專項“種禽場主要垂直傳播性疾病疫情調(diào)查”項目
【分類號】：S858.31

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