上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（临床研究） • 上一篇    下一篇

结核分枝杆菌Rv0315重组蛋白ELISPOT辅助诊断活动性肺结核的应用价值

周业成1, 陈翠翠2, 叶 娟3, 赵俊伟4, 玄松花3, 杨志荣1, 张舒林4   

  1. 1.四川大学 生命科学学院， 成都 610065； 2.郑州大学 第一附属医院， 郑州 450001； 3.河南中医学院 分子生物学教研室， 郑州 450008； 4.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:2013-07-28 发布日期:2013-08-22
  • 通讯作者: 张舒林, 电子信箱: shulinzhang@sjtu.edu.cn; 杨志荣, 电子信箱: bioyang@163.com。
  • 作者简介:周业成（1988—），男，硕士生; 电子信箱: zyc19880615@126.com。
  • 基金资助:

    上海市科委科技支撑项目（12441903300）；国家自然基金（81271794）；“十二五”国家科技重大专项（2013ZX10003002-005）

Diagnostic value of ELISPOT by recombinant Rv0315 protein of Mycobacterium tuberculosis in detection of active pulmonary tuberculosis

ZHOU Ye-cheng1, CHEN Cui-cui2, YE Juan3, ZHAO Jun-wei4, XUAN Song-hua3, YANG Zhi-rong1, ZHANG Shu-lin4   

  1. 1.College of Life Sciences, Sichuan University, Chengdu 610065, China; 2.The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450001, China; 3.Department of Biochemistry, Henan University of Traditional Chinese Medicine, Zhengzhou 450008, China； 4.Department of Medical Microbiology and Parasitology, Basic Medical College, Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2013-07-28 Published:2013-08-22
  • Supported by:

    Shanghai Science and Technology Committee Foundation, 12441903300; National Natural Science Foundation of China, 81271794; National Major Science and Technology Research Project in the “12th Five-year Plan”, 2013ZX10003002-005

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摘要：

目的 用酶联免疫斑点试验（ELISPOT）评价结核分枝杆菌重组蛋白Rv0315在活动性肺结核病诊断中的应用价值。方法 选取初治肺结核病患者25例和健康体检者28名分别作为结核组和对照组,应用Rv0315蛋白作为抗原对受试者外周血单核细胞进行ELISPOT检测斑点形成细胞（SFCs）的数量，判断结核分枝杆菌感染情况，并与结核早期分泌靶向抗原（ESAT6）和培养滤液蛋白10 （CFP10）抗原的检测结果进行比较。结果 以Rv0315、EAST6和CFP10作为抗原，用ELISPOT方法检测结核分枝杆菌感染的敏感度分别为84.0% （21/25）、80.0% （20/25）和92.0% （23/25），特异度分别为89.2% （25/28）、89.2% （25/28）和92.8% （26/28），其中Rv0315的敏感度高于EAST6但低于CFP10。结论 应用重组蛋白Rv0315作为抗原建立的ELISPOT方法具有应用于活动性肺结核病辅助诊断的潜在价值。

关键词: 结核分枝杆菌, Rv0315, 酶联免疫斑点技术, 细胞免疫

Abstract:

Objective To investigate the application of enzyme-linked immunospot assay (ELISPOT) by recombinant Rv0315 protein of Mycobacterium tuberculosis in the auxiliary diagnosis of active pulmonary tuberculosis. Methods A total of 25 patients with initial pulmonary tuberculosis (tuberculosis group) and 28 healthy people (control group) were enrolled. The peripheral blood mononuclear cells of all participants were co-cultured with Rv0315, earlier secreted antigen target 6 (EAST6) and culture filtrate protein 10 (CFP10) antigens, and spot forming cells (SFCs) were enumerated by ELISPOT. Results The values of sensitivity of Rv0315, EAST6 and CFP10 antigens by ELISPOT were 84.0% (21/25), 80.0% (20/25) and 92.0% (23/25) respectively, and those of specificity were 89.2% (25/28), 89.2% (25/28) and 92.8% (26/28) respectively. Rv0315 was more sensitive than EAST6, but was less sensitive in comparison with CFP10 in diagnosis of Mycobacterium tuberculosis infection. Conclusion ELISPOT with recombinant Rv0315 protein as antigen has the potential value in the auxiliary diagnosis of active pulmonary tuberculosis.

Key words: Mycobacterium tuberculosis, Rv0315, enzyme-linked immunospot assay, cellular immunity