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Development and application of an indirect ELISA for the detection of antibodies to novel duck reovirus

文献类型： 外文期刊

作者： Yun, Tao 1 ; Chen, Haipeng 2 ; Yu, Bin 1 ; Zhang, Cun 1 ; Chen, Liu 1 ; Ni, Zheng 1 ; Hua, Jionggang 1 ; Ye, Weicheng 1 ;

作者机构： 1.Zhejiang Acad Agr Sci, Inst Anim Husb & Vet Sci, Hangzhou 310021, Zhejiang, Peoples R China

2.Shandong Agr Univ, Coll Anim Sci & Vet Med, Tai An 271018, Shandong, Peoples R China

关键词： Novel duck reovirus (N-DRV);sigma C protein;Indirect ELISA;Serological survey

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN：

年卷期：

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收录情况： SCI

摘要： A novel duck reovirus (N-DRV) disease emerged in China in 2000 and it has become an epidemic genotype. A test for detection of virus-specific antibodies in serum samples would be useful for epidemiological investigations. Currently, Currently, serological assays for N-DRV diagnosis are not available. A test for detection of virus-specific antibodies in serum samples would be useful for epidemiological investigations. In this study, a highly sensitive and specific indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to N-DRV was developed. The outer capsid (sigma C) of N-DRV was cloned and expressed in Escherichia coil as a coating antigen. The antigen concentration and serum dilution were optimized using a checkerboard titration. Furthermore, the specificity of sigma C-ELISA assay was confirmed by cross checking with other duck viral pathogens. In comparison with the western blot, the sensitivity and specificity of the sigma C-ELISA was 92.6% and 88.9%, respectively, and agreement of two tests was excellent with kappa value of 0.786 (p < 0.05). A serological survey was performed using the assay on serum samples from different age and species of duck flocks in the Zhejiang and Jiangsu Province, China. The seropositive rate of the 1209 serum samples was 57.7%. In conclusion, the developed sigma C-ELISA assay is a very specific and sensitive test that will be useful for large-scale serological survey in N-DRV infection and monitoring antibodies titers against N-DRV. (C) 2015 Elsevier B.V. All rights reserved.

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