Multivalent nanobody-based sandwich enzyme-linked immunosorbent assay for sensitive detection of porcine reproductive and respiratory syndrome virus

文献类型： 外文期刊

第一作者： Sun, Mingxia

作者： Sun, Mingxia;Sun, Yue;Yang, Yongbo;Zhao, Man;Zhang, Minmin;Xia, Dasong;Wang, Tao;Gao, Yanfei;Wang, Shanghui;Wang, Haiwei;Cai, Xuehui;An, Tongqing;An, Tongqing;Cai, Xuehui;Cao, Dan;Cai, Xuehui;An, Tongqing

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关键词： PRRSV; Multivalent nanobody; Nanobody-HRP; Sandwich ELISA

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.2； 五年影响因子：7.8 ）

ISSN： 0141-8130

年卷期： 2024 年 258 卷

页码：

收录情况： SCI

摘要： The pandemic of the porcine reproductive and respiratory syndrome virus (PRRSV) has caused huge economic losses and continues to threaten the swine industry worldwide. Nucleocapsid protein (N protein) is the primary antigen of PRRSV for development of sensitive diagnostic assays. Two high affinity nanobodies against N protein, Nb12 and Nb35, were selected and employed to develop a sandwich ELISA. Further we improved the ELISA method to obtain greater sensitivity, a trivalent nanobody (3 x Nb35) and a bivalent nanobody-HRP fusion protein (2 x Nb12-HRP) were expressed and used. This modified ELISA was found to have high sensitivity for detecting PRRSV, with a detection limit of 10 TCID50/ml (median tissue culture infectious dose), which was approximately 200-fold greater than the single-copy nanobody-based sandwich ELISA. The developed assay shows high specificity and can detect almost all circulating lineages of PRRSV-2 in China. This study provides suggestions for reforming nanobodies and for the further development of multivalent nanobody-based ELISAs for other various viruses.

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