Development of an Epitope-Based Competitive ELISA for the Detection of Antibodies against Tibetan Peste des Petits Ruminants Virus

文献类型： 外文期刊

第一作者： Yu, Rui-Song

作者： Yu, Rui-Song;Zhu, Yu-Min;Dong, Shi-Juan;Li, Zhen;Zhang, Guo-Rui;Yu, Rui-Song;Zhu, Yu-Min;Dong, Shi-Juan;Li, Zhen;Zhang, Guo-Rui;Lei, Zhi-Hai;Zeng, Jiang-Yong;Zhu, Se;Duoji, Ciren;Dunzhu, Luobu

作者机构：

关键词： Competitive ELISA;Peste des petits ruminants virus;N gene;Epitope;Antibody

期刊名称：INTERVIROLOGY （ 影响因子：1.763； 五年影响因子：1.929 ）

ISSN： 0300-5526

年卷期： 2013 年 56 卷 1 期

页码：

收录情况： SCI

摘要： Aims: To develop an effective diagnostic kit, based on a competitive ELISA-based system (cELISA), for detecting serum antibody against peste des petits ruminants virus (PPRV). Methods: Epitope peptides of the nucleocapsid (N) protein of Tibetan PPRV were synthesized chemically and injected into rabbits to prepare hyperimmune antisera. Test sera were incubated simultaneously with hyperimmune antisera and added to the wells of ELISA plates coated previously with recombinant N protein. Horseradish peroxidase-conjugated goat anti-rabbit antibody was employed to detect the quantity of hyperimmune antisera combined with recombinant N protein. Results: A cELISA has been developed for monitoring PPRV infections with a cutoff value of 35. Relative sensitivity and specificity values of the epitope-based cELISA were 96.18 and 91.29%, respectively, when compared with a commercial cELISA kit in a test involving 1,039 serum samples. Conclusion: We report an efficient method for preparing antibody suitable for incorporation into a cELISA that can be used routinely for the detection of PPRV antibodies in serum samples. The method eliminated the requirement for virus culture and monoclonal antibody preparation, reduced the biorisk posed by virus-dependent manipulations, and the performance of the resultant cELISA compared favorably with a commercially available cELISA kit. Copyright (c) 2012 S. Karger AG, Basel

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