Functional epitope mapping of porcine reproductive and respiratory syndrome virus nsp7α and nsp7β using monoclonal antibodies and development of a feasible ELISA

文献类型： 外文期刊

第一作者： Han, Song

作者： Han, Song;Duan, Hongyong;Yang, Nan;Wang, Shumao;Gao, Fei;Jiang, Yifeng;Zhou, Yanjun;Tong, Guangzhi;Li, Liwei;Zhao, Kuan;Duan, Hongyong;Tang, Chaozhi;Li, Liwei

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关键词： Nonstructural protein 7; Antigenic epitope; Enzyme-linked immunosorbent assay

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.5； 五年影响因子：8.7 ）

ISSN： 0141-8130

年卷期： 2025 年 322 卷

页码：

收录情况： SCI

摘要： Porcine reproductive and respiratory syndrome virus (PRRSV) poses a significant threat owing to its considerable economic impact on the global swine industry. Nonstructural protein 7 (nsp7) plays an essential role in inducing the humoral immune response, resulting in the production of high and persistent antibodies in PRRSV-infected pigs. Therefore, nsp7 is considered the most appropriate marker for diagnostic development. Herein, we successfully expressed nsp7 using a prokaryotic expression system and generated eleven monoclonal antibodies. All antibodies exhibited specific reactivity and recognized the linear B-cell epitopes. By epitope mapping and antigenicity analysis, one novel epitope of nsp7 alpha (141IPLPPKVLE149) and three novel epitopes of nsp7 beta (152PNAWGDEDRL161, 184KFWDKNSGD192, 211GDPVDFDPET220) were further identified. These epitopes were conserved and located on the surface of the nsp7 spatial structure. PRRSV-infected pigs exhibited strong antibody responses specific for the identified epitopes. The enzyme-linked immunosorbent assay (ELISA) developed using two epitope peptides of nsp7 alpha and nsp7 beta showed high sensitivity, good specificity, and a 94.0 % coincidence rate with the commercial kit. The antibody was detected in pig sera using ELISA at 14 days post-infection, indicating its potential for PRRSV monitoring. The collective findings proved to be effective for detection and have promising application prospects for epidemiological surveillance and vaccine research.

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