数字农科院2.0

Epitope Identification and Application for Diagnosis of Duck Tembusu Virus Infections in Ducks

文献类型： 外文期刊

作者： Li, Chenxi;Shaozhou, Wulin;Bai, Xiaofei;Zhang, Qingshan;Hua, Ronghong;Liu, Ming;Zhang, Yun;Liu, Junyan;Liu, Jyung-Hurng

作者机构：

关键词： duck Tembusu virus;E protein epitopes;type specific and cross-reactive epitopes;E protein 3D structure;diagnosis

期刊名称： VIRUSES-BASEL

ISSN： 1999-4915

年卷期： 2016 年 8 卷 11 期

页码：

收录情况： SCI

摘要： Duck Tembusu virus (DTMUV) causes substantial egg drop disease. DTMUV was first identified in China and rapidly spread to Malaysia and Thailand. The antigenicity of the DTMUV E protein has not yet been characterized. Here, we investigated antigenic sites on the E protein using the non-neutralizing monoclonal antibodies (mAbs) 1F3 and 1A5. Two minimal epitopes were mapped to (LD)-L-221/NLPW225 and (87)YAEYI(91) by using phage display and mutagenesis. DTMUV-positive duck sera reacted with the epitopes, thus indicating the importance of the minimal amino acids of the epitopes for antibody-epitope binding. The performance of the dot blotting assay with the corresponding positive sera indicated that YAEYI was DTMUV type-specific, whereas (LD)-L-221/NLPW225 was a cross-reactive epitope for West Nile virus (WNV), dengue virus (DENV), and Japanese encephalitis virus (JEV) and corresponded to conserved and variable amino acid sequences among these strains. The structure model of the E protein revealed that YAEYI and LD/NLPW were located on domain (D) II, which confirmed that DII might contain a type-specific non-neutralizing epitope. The YAEYI epitope-based antigen demonstrated its diagnostic potential by reacting with high specificity to serum samples obtained from DTMUV-infected ducks. Based on these observations, a YAEYI-based serological test could be used for DTMUV surveillance and could differentiate DTMUV infections from JEV or WNV infections. These findings provide new insights into the organization of epitopes on flavivirus E proteins that might be valuable for the development of epitope-based serological diagnostic tests for DTMUV.

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