畜牧兽医学报2017，Vol.48Issue(10)：1958-1968,11.DOI:10.11843/j.issn.0366-6964.2017.10.019
鸭坦布苏病毒E蛋白特异性抗原表位鉴定及Epitope-ELISA血清学诊断方法建立
Unique Antigenic Epitope Identification of Duck Tembusu Virus Envelope Protein and Development of Epitope-ELISA
摘要
Abstract
In this study,a unique antigenic epitope 87YAEYI91 on E protein of duck Tembusu virus (DTMUV) was identified by phage display technology with purified monoclonal antibodies (MAb) 1A5 against DTMUV E protein.To determine whether the 87YAEYI91 sequence is conserved among the E proteins of flaviviruses,we aligned the E protein amino acid sequences of flaviviruses.The results revealed that YAEYI was highly conservative in DTMUV strain,but the epitope sequence wasn't conserved among the corresponding amino acid sequence of the E protein in other strains of flavivirus.Dot blotting assay showed that epitope 87 YAEYI91 had a good affinity with positive sera to DTMUV,and there was not cross-reacting with JEV-,DENV-,WNV-positive sera,suggesting that 87YAEYI91 was DTMUV unique epitope.An Epitope-ELISA assay was established by using unique epitope (87YAEYI91) fused protein and used to test duck serum samples which were also detected by the Neutralization Test as the standard method.According to results,the specificity of this test was 100％.The sensitivity of this epitope-based peptide serologic test for DTMUV infection was 96％.So,Epitope-ELISA was a specific method for detecting antibodies against DTMUV.关键词
鸭坦布苏病毒/抗原表位/交叉反应性/Epitope-ELISAKey words
duck Tembusu virus/epitope/cross-reacting/Epitope-ELISA分类
农业科技引用本文复制引用
李晨曦,白小飞,邵周伍林,张青山,刘明,张云..鸭坦布苏病毒E蛋白特异性抗原表位鉴定及Epitope-ELISA血清学诊断方法建立[J].畜牧兽医学报,2017,48(10):1958-1968,11.基金项目
十三五重点研发项目(2016YFD0500100) （2016YFD0500100）
国家自然科学基金面上项目(31670153) （31670153）
中国农业科学院协同创新项目(CAAS-XTCX2016011-04-9) （CAAS-XTCX2016011-04-9）
