利用核苷类逆转录酶抑制剂去除疫苗毒种中禽网状内皮组织增生症病毒污染

发布时间：2018-05-12 04:22

本文选题：禽网状内皮组织增生症病毒 + azidothymidine　；参考：《中国兽医学报》2017年06期

【摘要】：使用污染有禽网状内皮组织增生症病毒(REV)的弱毒疫苗被认为是REV流行的原因之一,去除疫苗毒种中REV污染是生产合格疫苗的基础环节,本研究试图利用核苷类逆转录酶抑制剂类药物Azidothymidine(LAM)和Lamivudine(AZT)去除疫苗毒种中REV污染。首先通过在培养液中添加不同质量浓度AZT或LAM连续传代以及利用CCK-8测定细胞活性的方法,证实了在培养液中添加不超过5mg/L的抑制剂类药物对于DF-1细胞复制和活性没有影响。体外试验证实,当在培养液中分别添加5mg/L的AZT或LAM以及两者联合应用时均可以显著抑制REV在DF-1细胞上的复制,通过在新城疫弱毒疫苗中人为添加REV模拟AZT或LAM以及两者联合应用对疫苗毒种中REV污染的净化作用,结果显示经药物干预4代后即将REV去除,并且净化后毒种检验合格。本研究证实了AZT或LAM在体外对REV的抑制作用,并在人工模拟试验中成功净化了疫苗毒种中污染的REV。
[Abstract]:The use of attenuated vaccine contaminated with avian reticuloendotheliosis virus (revv) is considered to be one of the reasons for the prevalence of REV. The removal of REV contamination from vaccine strains is the basic link in the production of qualified vaccines. The aim of this study was to remove REV contamination from vaccine strains by using nucleoside reverse transcriptase inhibitors Azidothymidine lam and Lamivudine AZT. Firstly, by adding different concentrations of AZT or LAM to the culture medium for continuous passage and using CCK-8 to determine the cell activity, it was proved that the addition of no more than 5mg/L inhibitor to the culture medium had no effect on the replication and activity of DF-1 cells. In vitro experiments showed that AZT or LAM of 5mg/L and their combination could significantly inhibit the replication of REV in DF-1 cells. By artificially adding REV to attenuated Newcastle disease vaccine, AZT or LAM were added to the vaccine, and the combined use of the two methods was used to purify the REV contamination in the vaccine. The results showed that REV would be removed after 4 generations of drug intervention, and the purified virus could be tested. This study confirmed the inhibitory effect of AZT or LAM on REV in vitro, and successfully purified the contaminated revs from vaccine strains in artificial simulation test.
【作者单位】：山东农业大学动物医学院;
【基金】：国家自然科学基金青年科学基金资助项目(31402226)
【分类号】：S852.65

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