禽弱毒疫苗中REV低剂量污染检测方法的比较

发布时间：2018-03-11 16:21

本文选题：禽网状内皮组织增生症病毒　切入点：污染　出处：《中国兽医学报》2017年06期 　论文类型：期刊论文

【摘要】：使用污染有禽网状内皮组织增生症病毒(REV)的弱毒疫苗是REV感染途径之一,而通常弱毒疫苗中REV的污染剂量较低,难以检测,本研究对不同方法在检测弱毒疫苗中REV低剂量污染进行了比较研究。将定量好的REV以不同剂量分别人为污染一批商品化新城疫病毒(NDV)弱毒疫苗,提取核酸后分别以TaqMan探针荧光定量RT-PCR检测、常规RT-PCR检测以及PCR产物结合核酸斑点杂交检测等不同方法进行检测。结果显示常规RTPCR检测仅可检测到10TCID50/羽份的REV污染,而另外两种方法均检测到1TCID50/羽份的低剂量REV污染。将人为污染1TCID50/羽份和2TCID50/羽份REV的NDV弱毒疫苗各接种10只SPF鸡,定期针对REV抗体及其核酸进行检测,结果显示在免疫污染疫苗后7周内REV抗体全部为阴性,而核酸检测均可以检测到一定比例的阳性,提示通过接种SPF鸡检测弱毒疫苗中REV污染时,仅仅通过抗体判定可能会对一些剂量较低的REV污染造成漏检,通过结合对病原的检测有助于降低漏检率。
[Abstract]:The use of attenuated vaccines contaminated with avian reticuloendotheliosis virus (revv) is one of the routes of REV infection, and REV contamination in attenuated vaccines is usually low and difficult to detect. In this study, the low dose pollution of REV in attenuated vaccine by different methods was compared. A batch of commercial Newcastle disease virus (NDV) attenuated vaccine was contaminated with different doses of REV. The nucleic acid was extracted and detected by TaqMan probe fluorescence quantitative RT-PCR detection, conventional RT-PCR detection and PCR product combined with nucleic acid dot hybridization respectively. The results showed that only 10TCID 50 / plume REV contamination could be detected by conventional RTPCR detection. The other two methods detected low dose REV contamination of 1TCID50 / plume. Ten SPF chickens were inoculated with the NDV attenuated vaccine of 1TCID50 / plume and 2TCID50 / plume REV respectively, and the antibody to REV and its nucleic acid were detected regularly. The results showed that all the REV antibodies were negative within 7 weeks after immunization, but the nucleic acid detection could detect a certain proportion of positive, suggesting that the REV contamination in the attenuated vaccine could be detected by inoculation of SPF chickens. The detection of REV with low dose may be caused by the detection of antibodies, and the detection of pathogens can help to reduce the rate of missed detection.
【作者单位】：山东农业大学动物医学院;
【分类号】：S852.65

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