上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (11): 1426-.doi: 10.3969/j.issn.1674-8115.2012.11.007

• 专题报道（病原微生物学） • 上一篇    下一篇

结核分枝杆菌特异性抗原ESXO的重组制备及其免疫原性评估

封 莉1, 叶 娟2, 王凤平1, 赵俊伟3, 吴兴福1, 张舒林3   

  1. 1.江苏省苏州市第五人民医院检验科, 苏州 215007； 2.河南中医学院分子生物学教研室, 郑州 450008； 3.上海交通大学 基础医学院病原生物学教研室, 上海 200025
  • 出版日期:2012-11-28 发布日期:2012-11-30
  • 作者简介:封 莉（1976—）, 女, 主管检验师, 学士；电子信箱: xfsh361@163.com。
  • 基金资助:

    国家自然基金科学（81271794）；上海市科委科技支撑项目（12441903300）

Recombination, preparation and immunological evaluation of Mycobacterium tuberculosis specific antigen ESXO

FENG Li1, YE Juan2, WANG Feng-ping1, ZHAO Jun-wei3, WU Xing-fu1, ZHANG Shu-lin3   

  1. 1.Suzhou No.5 People's Hospital, Suzhou 215007, China；2.Department of Biochemistry, Henan University of Traditional Chinese Medicine, Zhengzhou 450008, China；3.Department of Microbiology and Parasitology, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2012-11-28 Published:2012-11-30
  • Supported by:

    National Natural Science Foundation of China, 81271794；Shanghai Science and Technology Committee Foundation, 12441903300

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摘要：

目的 评估结核分枝杆菌（MTB）重组特异性抗原ESXO在大肠埃希菌中的高效表达，并评价其免疫原性。方法 采用常规分子克隆方法获得MTB重组蛋白ESXO。酶联免疫吸附试验(ELISA)方法检测84份确诊结核病（TB）患者血清和48份健康人血清中相应的抗结核ESXO抗体水平，评价血清学抗原活性，并与结核早期分泌蛋白ESAT-6和CFP-10的检测结果进行比较。结果 重组特异性抗原ESXO在大肠杆菌中获得成功表达，其在E.coli BL 21 plysS (DE3)中以可溶性和包涵体两种形式存在，相对分子质量（27 300）与预期相符，血清学抗原活性评估结果显示其特异度和敏感度分别为94.0%(45/48)和32.1%(27/84)，特异度与ESAT-6和CFP-10相当，敏感度高于ESAT-6。结论 结核特异性抗原ESXO有望成为新的TB诊断与疫苗设计的候选抗原。

关键词: 分枝杆菌, 结核, 克隆, ESXO, 基因表达, 血清学诊断

Abstract:

Objective To determine the expression of recombinant Mycobacterium tuberculosis (MTB) specific antigen ESXO in E.coli, and evaluate its immunogenicity. Methods Recombinant ESXO protein of MTB was obtained with conventional molecular cloning method. The serum antibody against ESXO was detected by enzyme linked immunosorbent assay (ELISA) in 84 patients with tuberculosis (TB) and 48 healthy controls, the serum immunogenicity was evaluated, and the results were compared with findings in detection of ESAT-6 and CFP-10 protein for serodiagnosis of TB. Results The recombinant specific antigen ESXO was expressed as soluble and inclusion body protein in E.coli BL21 plysS (DE3), with the relative molecular weight of 27 300, which was in line with expectations. The immunogenic evaluation of recombinant ESXO protein indicated that its specificity and sensitivity reached 94.0% (45/48) and 32.1% (27/84) respectively, which was equal to ESAT-6 and CFP-10 in specificity and higher than ESAT-6 in sensitivity. Conclusion The specific antigen ESXO is expected to be a novel candidate antigen for the diagnosis and vaccine design in TB.

Key words: Mycobacterium tuberculosis, clone, ESXO, gene expression, serodiagnosis