Characterization of the sigma B-encoding genes of muscovy duck reovirus: sigma C-sigma B-ELISA for antibodies against duck reovirus in ducks

文献类型： 外文期刊

第一作者： Zhang, Yun

作者： Zhang, Yun;Guo, Dongchun;Liu, Ming;Geng, Hongwei;Hu, Qilin;Liu, Yin;Liu, Nihong

作者机构：

关键词： muscovy duck reovirus;sigma B/sigma C-encoding gene;cloning;expression;sigma C-sigma B-ELISA

期刊名称：VETERINARY MICROBIOLOGY （ 影响因子：3.293； 五年影响因子：3.599 ）

ISSN： 0378-1135

年卷期： 2007 年 121 卷 3-4 期

页码：

收录情况： SCI

摘要： The (sigma B/sigma C-encoding genes of muscovy duck reovirus (DRV) S12 strain were cloned, sequenced, and expressed in Escherichia coli. The (sigma C-encoding gene of DRV showed only 21-22% identity to that of avian reovirus (ARV) at both nucleotide and amino acid level. The sigma B-encoding gene of DRV comprised 1163 bp with one open reading frame (ORF). The ORF comprised 1104 bp and encoded 367 amino acids with a predicted molecular mass of 40.44 kDa. A zinc-binding motif and a basic amino acid motif were found within the predicted amino acid sequence of sigma B. The identities between the S12 and ARV were 59.3-64.0% and 60.9-62.5%, respectively, at the nucleotide and deduced amino acid levels. Phylogenetic analysis of the (sigma B-encoding gene sequence indicated that S12 separated as a distinct virus relative to other avian strains. The expressed sigma B/sigma C fusion proteins in E. coli could be detected, approximately 45 and 50 kDa, respectively, by duck anti-reovirus polyclonal serum. In addition, an ELISA (sigma B-sigma C-ELISA) using the expressed sigma B-sigma C proteins as coating antigen for detection of antibodies to DRV in ducks was developed. In comparison with the virus neutralization test and agar gel immuno-diffusion test (AGID), the (sigma B-sigma C-ELISA showed perfect specificity and sensitivity. The (sigma B-sigma C-ELISA did not react with the antisera to other duck pathogens, implying that these two proteins were specific in recognition of DRV antibodies. Taken together, the results demonstrated that (sigma B-sigma C-ELISA was a sensitive and accurate method for detecting antibodies to DRV. (c) 2006 Elsevier B.V. All rights reserved.

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