摘要
目的探讨胸腔积液PEMC中CD4＋T淋巴细胞分泌或产生IFN—γ的检测对结核性胸膜炎的辅助诊断价值。方法分离40例结核性胸腔积液（结核组）和30例恶性胸腔积液患者（疾病对照组）PELVIC，经克隆表达的早期分泌抗原靶6（ESAT-6）和培养滤液蛋白10（CFP-10）融合蛋白（简称“E／C”）刺激后，用ELISpot检测分泌IFN—γ细胞的斑点形成细胞（SFC）数量和流式细胞术（FCM）结合胞内细胞因子染色检测产生IFN-γ细胞比率，并比较这2项指标在结核组和疾病对照组间的差异，同时评价2种方法检测IFN-γ对结核性胸膜炎的辅助诊断效能。结果PEMC经E／C刺激后用ELISpot检测结核组SFC数量为205（125-450）SFC／5×10^4PEMC，疾病对照组为5（2～18）SFC／5×10^4PEMC，两组间的差异有统计学意义（u=20．00，P〈0．01）；FCM检测结核组CD4＋T淋巴细胞产生1FN—Y细胞比率为3．27％（1．81％-7．34％），疾病对照组为0．12％（0．06％-0．46％），两组问的差异有统计学意义（U=45．00，P〈0．01）。ELISpot检测PEMC经E／C刺激后分泌IFN-γ的敏感度为92．5％（37／40）、特异度为80．0％（24／30）、阳性预测值为0．86、阴性预测值为0．89、阳性似然比为4．63、阴性似然比为0．09和准确度为87．1％；FCM检测经E／C刺激后产生IFN-γ的敏感度为87．5％（35／40）、特异度为90．0％（27／30）、阳性预测值为0．92、阴性预测值为0．84、阳性似然比为8．75、阴性似然比为o．14和准确度为88．6％。结论经E／C刺激后，用FCM和ELISpot检测CD4＋T淋巴细胞产生和分泌IFN—γ细胞的方法对结核性胸膜炎诊断的敏感度和特异度高，且对结核性胸膜炎有辅助诊断价值。
Objective To explore the value of IFN- γ produced or secreted by CD4＋ T Lymphocytes from pleural effusion mononuclear ceils for the diagnosis of tuberculous pleurisy （plTB） . Methods The PEMCs of 40 patients with tuberculous pleural effusion and 30 patients with malignancy pleural effusion were selected as the tuberculosis and disease control groups, then co-cultured with the early secretory antigenic target 6（ESAT-6） and culture filtered protein 10 （CFP-10） fusion protein （E/C）. The numbers of spot forming ceils （SFC） secreting IFN-γ were enumerated by ELISpot and the ratios of cells producing IFN-γ were detected by flow cytometry and intracellular cytokine staining. Moreover, the two indicators were compared between tuberculosis and disease control groups to evaluate the 2 methods detecting IFN-γ in the diagnosis of plTB. Results After E/C stimulation, the numbers of SFC were 205 （125-450） SFC/5 × 10^4 PEMC in tuberculosis group and 5 （2-18） SFC/5 × 10^4 PEMC in disease control group by ELISpot. The difference between two groups was statistically significant （U = 20.00, P 〈0.01）. The proportion of IFN- γ -secreting CD4＋ T lymphocytes was 3.27% （1.81% - 7.34%） in tuberculosis group and 0.12% （0.06% - 0.46%） in control group detected by FCM. The difference between the two groups was statistically significant （U = 45.00, P 〈0.01）. The indicators of ELISpot in detection of IFN-γ which was secreted by PEMC after co-cultured with E/C were as follows： sensitivity 92.5% （37/40）, specificity 80.0% （24/30）, positive predictive value 0.86, negative predictive value 0.89, positive likelihood ratio 4.63, negative likelihood ratio 0.09 and accuracy 87.1%; and for FCM, they were 87.5% （35/40）, 90.0% （27/30）, 0.92, 0.84, 8.75 and 0.14, respectively and accuracy 88.6%. Coneluslon After E/C stimulation, the assay for IFN- γ -secreting CD4＋ T Lymphocytes by FCM and ELISpot is highly sensitive and specific for diagnosis of plTB as an auxiliary method.
出处
《结核病与胸部肿瘤》
2011年第3期166-171,共6页
Tuberculosis and Thoracic Tumor
基金
国家自然科学基金资助项目（30771916）
国家重大专项“结核病诊断分子标识研究”2008ZX10003-005）
关键词
胸膜
结核
免疫酶技术
流式细胞术
Tuberculosis, pleural
Inmmnoenzyme techniques
Flow cytometry
