Development and validation of a DIVA ELISA for differentiating BEFV infected from vaccinated animals

文献类型： 外文期刊

第一作者： He, Chenxiang

作者： He, Chenxiang;Gao, Shandian;Du, Junzheng;Tian, Zhancheng;Guan, Guiquan;Yin, Hong;Yin, Hong

作者机构：

关键词： Bovine ephemeral fever virus; Differentiation of Infected from Vaccinated Animals (DIVA); ELISA

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.623； 五年影响因子：2.239 ）

ISSN： 0166-0934

年卷期： 2022 年 310 卷

页码：

收录情况： SCI

摘要： Inactivated vaccine is considered safe and used for prevention of bovine ephemeral fever in several endemic countries. To differentiate between BEFV-infected and vaccinated animals, we developed an ELISA capable of detecting infection-related antibodies against BEFV. Recombinant proteins, including N, P, M, L, G(NS), alpha(2), beta and gamma, were expressed in E. coli and screened by Western blotting and ELISA. The results showed G(NS), alpha(2) and beta specifically reacted with sera from BEFV infected cattle but not sera from vaccinated cattle. A DIVA ELISA based on a C-terminal truncated form of G(NS) was developed, with 100% sensitivity and 98.0% specificity at a sample to positive-control optical density ratio (S/P) threshold of 0.18. Specificity analysis showed that the assay has no cross-reactivity with antisera of other common bovine viruses. Anti-G(NS) antibody appears at 3-4 days post infection (dpi) and persists up to 240-300 dpi in the experimentally infected cattle. Sero-epidemiological survey using sera collected from vaccinated cattle in an endemic area in Jiangsu Province revealed sero-positive rate of 2.36% (6/254), indicating that the DIVA ELISA could be used as a reliable diagnostic tool for differentiating BEFV infected from vaccinated animals.

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