Enhanced expression of the E-rns protein of classical swine fever virus in yeast and its application in an indirect enzyme-linked immunosorbent assay for antibody differentiation of infected from vaccinated animals

文献类型： 外文期刊

第一作者： Luo, Yuzi

作者： Luo, Yuzi;Li, Lin;Dong, Mei;Xu, Jingjing;Shao, Lina;Lei, Jianlin;Li, Na;He, Wen-Rui;Zhao, Bibo;Li, Su;Li, Yongfeng;Sun, Yuan;Qiu, Hua-Ji;Austermann-Busch, Sophia;Becher, Paul;Liu, Lihong;Luo, Yuzi;Li, Lin;Xu, Jingjing;Shao, Lina;Lei, Jianlin;He, Wen-Rui;Zhao, Bibo;Li, Su;Li, Yongfeng;Liu, Lihong;Sun, Yuan;Qiu, Hua-Ji

作者机构：

关键词： Classical swine fever virus;E-rns protein;Yeast expression;Indirect enzyme-linked immunosorbent assay;Differentiation of infected from vaccinated animals (DIVA)

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：

页码：

收录情况： SCI

摘要： Classical swine fever (CSF), caused by classical swine fever virus (CSFV), is a devastating disease of swine worldwide. Although a mandatory vaccination with the modified live vaccine C-strain has been implemented in China for decades, CSF remains a serious threat to the swine industry. To facilitate the control and eradication of CSF in China, the E2-based marker vaccine rAdV-SFV-E2, an adenovirus-delivered, alphavirus replicon-vectored vaccine, has been developed. Accordingly, an accompanying discriminatory test that allows differentiating infected from vaccinated animals (DIVA) is required. Here, the enhanced expression of E-rns protein of CSFV was achieved in the methyltropic yeast Pichia pastoris by codon-optimization of the E-rns gene, and an indirect enzyme-linked immunosorbent assay (iELISA) based on the yeast-expressed E-rns (yE(rns)) was developed and evaluated. The optimized iELISA was able to detect CSFV-specific antibodies in the serum samples from the CSFV-infected pigs as early as 6 days post-infection, and discriminate the CSFV-infected pigs from those vaccinated with rAdV-SFV-E2. The iELISA was evaluated using a panel of swine sera, and showed comparable sensitivity (94.6%) and specificity (97.1%), and the consistence rates with the virus neutralization test were 96.8% for CSFV-infected swine sera, 83.3% for C-strain-vaccinated swine sera, and 95.0% for field swine sera. In addition, the iELISA showed higher sensitivity (90.4%) compared with PrioCHECK CSFV E-rns (59.6%). Taken together, the yE(rns)-based iELISA is specific and sensitive, representing a promising DIVA test for E2-based marker vaccines against CSF. (C) 2015 Elsevier B.V. All rights reserved.

分类号： R37

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