流式细胞术检测产IFN-γ的T淋巴细胞及在肺结核诊断中的应用  

The application of detection of IFN-γ-secreting T lymphocytes by flow cytometry in diagnosis of pulmonary tuberculosis

作　　者：田华 武尔杰 陆仁飞[3] 罗文明[3] 袁佳璐 顾德林 

机构地区：[1]南通市第六人民医院检验科,江苏226011 [2]南通产源生物科技有限公司 [3]南通市第三人民医院检验科

出　　处：《交通医学》2016年第3期227-230,共4页Medical Journal of Communications

基　　金：南通市社会事业科技创新与示范项目(HS149057)

摘　　要：目的:建立流式细胞术(flow cytometry,FCM)检测产IFN-γCD4^+T淋巴细胞的方法,探讨该方法对肺结核辅助诊断的应用价值。方法:收集42例肺结核患者及30例正常体检者肝素钠抗凝血,分离外周血单个核细胞,经结核杆菌特异性ESAT-6和CFP-10混合抗原刺激后,以流式细胞术检测经荧光抗体染色的产IFN-γ的CD4^+T淋巴细胞比率,以受试者工作曲线评价该方法对肺结核的诊断效能并与T-SPOT.TB法进行比较。结果:FCM法肺结核组产生IFN-γCD4^+T淋巴细胞数比率为(3.96±2.32)%,显著高于对照组的(0.62±0.26)%,差异有统计学意义(P<0.001)。FCM法受试者工作曲线下面积为0.9821,以阳性细胞比率0.93%为临界值,其诊断肺结核的灵敏度为95.24%,特异性为93.33%,与T-SPOT.TB法相近。结论:流式细胞术检测产IFN-γCD4^+T淋巴细胞的方法对肺结核具有一定的辅助诊断价值。Objective： To establish a detected method of IFN-γ-secreting CD4^＋T lymphocytes by flow cytometry（FCM） and explore the application of the method in diagnosis of pulmonary tuberculosis（PTB）. Methods： The peripheral blood mononuclear cells（PBMCs） of 42 patients with PTB and 30 normals receiving health examination were collected and co-cultured with the early secretory antigenic target 6（ESAT-6） and culture filtered protein 10（CFP-10） with fusion protein.The IFN-γ-secreting CD4^＋T lymphocytes were stained by immunofluorescence and detected by FCM. The diagnostic efficacy of the FCM method in PTB was evaluated by the receiver operating characteristic curve（ROC） and compared with the T-SPOT.TB method. Results： The ratio of IFN-γ-secreting CD4^＋T lymphocytes detected by FCM was 3.96%±2.32%in PTB, significantly higher than 0.62%±0.26% in the normal control（P〈0.001）. The area under ROC of the ratio by FCM was 0.9821. Take the radio 0.93% for the cut-off value to diagnosise PTB, the sensitivity and specificity were 95.24% and93.33% respectively. Those diagnostic data of FCM were all similar to T-SPOT.TB. Conclusions： The assay for IFN-γ-secreting CD4^＋T lymphocytes by FCM has subsidiary value for diagnosis of PTB.

关 键 词：流式细胞术 产γ-干扰素T淋巴细胞 T-SPOT.TB 肺结核 

分 类 号：R521[医药卫生—内科学]