不同抗原酶联免疫斑点检测技术对结核性胸膜炎辅助诊断价值的比较  

The comparison of different antigens in enzyme-linked immunospot assay for the diagnosis of tuberculous pleurisy

作　　者：杜凤娇[1] 陈曦[1] 刘菲[2] 刘洋[1] 曹廷明[1] 刘忠泉[1] 贾红彦[1] 邢爱英[1] 杜博平 古淑香[1] 马玙[1] 张宗德[1] 

机构地区：[1]北京市结核病胸部肿瘤研究所结核病分子生物学研究室,101149 [2]首都医科大学附属北京胸科医院结核一科

出　　处：《北京医学》2011年第12期981-985,共5页Beijing Medical Journal

基　　金：国家自然科学基金(30771916);艾滋病和病毒性肝炎等重大传染病防治科技重大专项(2008ZX10003-005)

摘　　要：目的探讨结核分枝杆菌不同抗原的酶联免疫斑点检测技术(ELISPOT)对结核性胸膜炎的辅助诊断价值。方法分离55例结核性胸膜炎患者和43例恶性胸腔积液患者的胸腔积液单个核细胞(PEMCs),经早期分泌抗原靶6kD(ESAT-6)、培养滤液蛋白10kD(CFP-10)、ESAT-6/CFP-10融合蛋白(E/C)、Rv3873和Rv3879c5个蛋白抗原共同孵育,进行ELISPOT试验,检测经抗原刺激后分泌干扰素-γ(IFN-γ)的效应T淋巴细胞(即斑点形成细胞,SFC)的数量,以中位数(四分位间距)表示。结果结核性胸膜炎组ESAT-6-ELISPOT、E/C-ELISPOT、CFP-10-ELISPOT、Rv3873-ELISPOT和Rv3879c-ELISPOT每孔的SFC数量分别为27(13～65)/5×104,26(12～52)/5×104,34(18～79)/5×104,26(8～50)/5×104和11(1～24)/5×104,均显著高于恶性胸膜积液组[1(0～2)/5×104,2(0～4)/5×104,2(1～4)/5×104,3(1～6)/5×104和1(0～3)/5×104,P均<0.01]。5个抗原中E/C的敏感性最高(92.7%),ESAT-6和Rv3879c的特异性最高(均为93.0%),ESAT-6的诊断准确性最高(91.8%)。结论 ESAT-6联合新抗原Rv3879c的ELISPOT能够辅助诊断结核性胸膜炎,但其特异性易受结核分枝杆菌潜伏感染的影响。Objective To explore different antigens of Mycobacterium tuberculosis （M.tb） in enzyme-linked immunospot assay （ELISPOT） for the auxiliary diagnosis of tuberculous pleurisy （plTB）. Methods The pleural effusion mononuclear cells （PEMCs） were separated in 55 patients of plTB and 43 patients with malignancy pleura] effusion （maPE）. Then 5 antigens were co-cultured： early secretory antigenic target 6 （ESAT-6）, culture filtered protein 10 （CFP-10）, ESAT-6/CFP-10 fusion protein （E/C）, Rv3873 and Rv3879c. Then T lymphocytes releasing interferon-γ/were detected （also spot forming cells, SFC） with a median （interquartile range） by ELISPOT. Results ESAT-6-ELISPOT, E/C-ELISPOT,CFP-10-ELISPOT,Rv3873-ELISPOT and Rv3879c-ELISPOT showed significantly higher SFC per well in plTB group [27（13-65）/5×10^4,26（12-52）/5×10^4,34（18-79）/5×10^4, 26（8-50）/5×10^4 and 11（1-24）/5×10^4 respectively] as compared to the maPE group [1（0-2）/5×10^4, 2（0-4）/5×10^4, 2（1-4）/5×10^4, 3（1-6）/5×10^4 and 1（0-3）/5×10^4 respectively, all P 〈 0.01]. E / C had the highest sensitivity （92.7%）. ESAT-6 and Rv3879c had the highest specificity （93.0）%. ESAT-6 had the highest diagnostic accuracy（91.8%） in the five antigens. Conclusion ESAT-6 combining with the novel antigen Rv3879c ELISPOT is a promising approach for the auxiliary diagnosis of plTB, but its specificity could be affected by latent tuberculosis infection.

关 键 词：结核分枝杆菌 酶联免疫斑点检测技术 胸腔积液 

分 类 号：R521.7[医药卫生—内科学]