南方医科大学学报2012，Vol.32Issue(11)：1667-1670,1679,5.DOI:10.3969/j.issn.1673-4254.2012.11.033
登革2型病毒特异性抗原片段的筛选及验证
Screening and identification of dengue virus type 2-specific antigens
摘要
Abstract
Objective To screen and identify dengue virus type 2 specific antigens and establish an enzyme-linked immunosorbent assay (ELBA) for detecting dengue virus type 2 antibody. Methods Using the bioinformatic software DNAstar and ANTHEPROT, we analyzed the hydrophilicity, flexibility, surface probability and antigenicity of dengue virus type 1-4, Japanese encephalitis virus, and Yellow fever virus M and E protein amino acid sequences, and also evaluated the influence of secondary structure. The specific epitopes of dengue virus type 2 were predicted according to the epitope location and amino acid sequence similarity, and the epitope conservation was assessed using the sequence information of different dengue virus type 2 strains in GenBank. Based on the results of bioinformatic analysis, 5 specific epitopes were amplified and inserted into the prokaryotic expression vector pET32a, which were transferred into E. coli Rosetta (DE3) for expression of the proteins. SDS-PAGE and Western blotting were used to identify the expressed proteins and test their antigenicities. The antigen selected by Western blotting was used to establish the ELISA system for dengue virus type 2 antibody detection. Results Bioinformatic analysis predicted 8 possible dengue virus type 2 specific epitopes, and 6 of them were efficiently expressed in E. coli. Western blotting confirmed 1 dengue virus type 2 specific antigen, the ELISA system for dengue virus antibody detection was successfully established using this specific antigen. Conclusion We have obtained a dengue virus type 2 specific antigen and established an ELISA system for detection of dengue virus type 2 antibody.关键词
登革2型病毒/抗原表位/原核表达/ELISAKey words
dengue virus type 2/ protein epitopes/ prokaryotic expression/ enzyme-linked immunosorbent assay分类
医药卫生引用本文复制引用
任瑞文,唐博恒,张培,胡文龙,洪文艳,刘建伟..登革2型病毒特异性抗原片段的筛选及验证[J].南方医科大学学报,2012,32(11):1667-1670,1679,5.基金项目
广东省科技计划项目(2010B031600125,2011B031500011) （2010B031600125,2011B031500011）
广州市科技计划项目(2010Y1-C151) （2010Y1-C151）
