数字农科院2.0

An Expeditious Neutralization Assay for Porcine Reproductive and Respiratory Syndrome Virus Based on a Recombinant Virus Expressing Green Fluorescent Protein

文献类型： 外文期刊

作者： Wang, Juan;Yan, Jiecong;Wang, Shuaiyong;Chen, Ronglin;Xing, Yanru;Liu, Qingyan;Gao, Shuolei;Zhu, Yuxiang;Li, Jiannan;Zhou, Yanjun;Shan, Tongling;Tong, Wu;Zheng, Hao;Kong, Ning;Jiang, Yifeng;Liu, Changlong;Tong, Guangzhi;Yu, Hai

作者机构：

关键词： PRRSV;infectious clones;reporter virus;enhanced GFP;neutralizing antibodies;neutralization assay

期刊名称： CURRENT ISSUES IN MOLECULAR BIOLOGY

ISSN： 1467-3037

年卷期： 2024 年 46 卷 2 期

页码：

收录情况： SCIE（2024版）

摘要： Due to the extensive genetic and antigenic variation in Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), as well as its rapid mutability and evolution, PRRS prevention and control can be challenging. An expeditious and sensitive neutralization assay for PRRSV is presented to monitor neutralizing antibodies (NAbs) in serum during vaccine research. Here, a PRRSV expressing eGFP was successfully rescued with reverse genetics based on the infectious clone HuN4-F112-eGFP which we constructed. The fluorescent protein expressions of the reporter viruses remained stable for at least five passages. Based on this reporter virus, the neutralization assay can be easily used to evaluate the level of NAbs by counting cells with green fluorescence. Compared with the classical CPE assay, the newly developed assay increases sensitivity by one- to four-fold at the early antibody response stage, thus saving 2 days of assay waiting time. By using this assay to unveil the dynamics of neutralizing antibodies against PRRSV, priming immunity through either a single virulent challenge or only vaccination could produce limited NAbs, but re-infection with PRRSV would induce a faster and stronger NAb response. Overall, the novel HuN4-F112-eGFP-based neutralization assay holds the potential to provide a highly efficient platform for evaluating the next generation of PRRS vaccines.

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