结核分枝杆菌抗原模拟肽的筛选和鉴定  

Screening and identification of the mimic peptide of Mycobacterium tuberculosis antigen

作　　者：孙文霞[1] 袁仕善[1] 谭云洪 张小萍 余艳艳 

机构地区：[1]湖南师范大学医学院分子生物学研究室,长沙410013 [2]湖南结核病防治研究所

出　　处：《中华微生物学和免疫学杂志》2012年第1期16-19,共4页Chinese Journal of Microbiology and Immunology

基　　金：湖南省高等学校科学研究项目（10C0920）;湖南省研究生科研创新项目（CX2010821）

摘　　要：目的应用噬菌体随机12肽库免疫淘筛结核分枝杆菌抗原模拟肽。方法以结核患者血清IgG为靶分子，3轮淘筛噬菌体随机12肽库，ELISA鉴定阳性噬菌体克隆，并进行测序分析。选择高频出现的阳性克隆用ELISA法初步分析其诊断能力。结果经3轮免疫筛选，噬菌体得到有效富集，12个阳性噬菌体克隆经测序分析获得6种短肽序列。高频出现的两个阳性克隆诊断结核病的敏感性分别为71．4％（A2）和55．4％（A7）。结论结核患者血清IgG筛选噬菌体随机12肽库获得了能结合抗结核抗体的噬菌体展示短肽。Objective To immunosereen the mimic peptides of Mycobaeterium tuberculosis antigen from phage displayed 12-mer peptide library. Methods Specific IgG was purified from sera of patients with TB and used as the target to immunoscreen a phage random peptide library of 12 amino acids. Positive clones which were obtained after three rounds of biopanning were detected by ELISA and sequenced. The diagnostic value of the high frequent positive clones were observed by ELISA. Results After 3 rounds of immunoscreening, the eluted phages were enriched effectively. Six kinds of animo acid sequence were obtained from twelve positive phage clones. Sensitivity of the two high frequent positive clones were 71.4% （ A2 ） and 55.4% （A7） respectively. Conclusion The antigen-mimic peptide was successfully screened from 12 random phage peptide library and the peptides can be recognized by tuberculosis patients＇ polyclonal antibodies.

关 键 词：结核分枝杆菌 噬菌体随机肽库 模拟肽 

分 类 号：R392[医药卫生—免疫学]