CFP-10/ESAT-6特异性IFN-g释放反应诊断活动性肺结核的研究  

The Study of ESAT-6/CFP-10 Fusion Protein Specific IFN-g Release Assay on Diagnosis of Active tuberculosis

作　　者：陈颖钰[1,2] 詹枝华[3] 郭爱珍[1,2] 项杰[3] 邓铨涛[1,2] 曾庆志 杜义祥 周金海[3] 魏武[5] 童庆伟[5] 陈焕春[1,2] 

机构地区：[1]华中农业大学农业微生物学国家重点实验室,武汉430070 [2]华中农业大学动物医学院预防兽医学省重点实验室,武汉430070 [3]武汉市结核病医院,武汉430083 [4]武汉市结核病防治所,武汉430030 [5]华中农业大学校医院,武汉430070

出　　处：《生物技术通报》2008年第S1期176-180,共5页Biotechnology Bulletin

基　　金：国家973项目(项目编号2006CB504400);国家"十一五"科技攻关奶业专项(项目编号:2006BAD04A05 2006BAD04A12);武汉市科技攻关课题(项目编号:20066002056)

摘　　要：前期研究已经建立了基于结核分枝杆菌特异性抗原CFP-10/ESAT-6融合蛋白刺激外周血单核细胞IFN-γ释放反应,本研究旨在证实该方法在活动性肺结核及结核菌潜伏感染诊断中的意义。实验对象包括111例当地医院收治的活动性肺结核病人(病例组)及283例某大学入学新生(健康对照者)。采集肝素抗凝血,分别加入含结核菌抗原CFP-10/ESAT-6、植物血凝素及无刺激物(PBS)的细胞培养孔中,培养过夜,次日收集血清,进行IFN-γ检测。同时,对其中58位病人志愿者及46位健康对照志愿者进行了结核菌素(PPD)皮内变态反应。病例组与健康对照组的PPD皮内变态反应阳性率分别为79.3%(46/58)和76.1%(35/46),无显著差异(P>0.05),表明PPD皮内变态反应不能用于活动性肺结核的检测。病例组IFN-γ体外释放反应的阳性率为95.5%(106/111),而健康对照组阳性率为16.3%(46/283),两组间均存在极显著差异,表明IFN-γ体外释放反应诊断活动性肺结核具有很高的灵敏度(95.5%)与良好的特异性。在健康组中,IFN-γ体外释放反应与PPD皮内变态反应的总符合率为50.0%,且IFN-g体外释放反应阳性者中,83.3%为PPD皮内变态反应阳性;在病例组中,二者的总符合率为72. 4%,IFN-g体外释放反应阳性者中,77.8%为PPD皮内变态反应阳性。CFP-10/ESAT-6特异性IFN-γ体外释放反应诊断活动性肺结核具有很高的灵敏度与特异性,PPD皮内变态反应由于受卡介苗免疫等因素影响在我国不能用于诊断活动性肺结核病与结核菌感染。Objective:To study the application of CFP-10/ESAT-6 specific IFN-γrelease assay previously established in diagnosis of active tuberculosis.Methods The heparized blood was collected from 111 active tuberculosis patients from the local hospital(patient group)and 283 healthy freshmen from some local university(control group)was collected and stimulated separately with CFP-10/ESAT-6 antigen,phytohemagglutinin(PHA)and saline overnight.The release of IFN-γin the culture supernatant was tested with a sandwich ELISA.Meanwhile,58 patient and 46 healthy control volunteers were subjected to PPD tuberculin skin test.Result The positive rates of PPD skin test for patient and control groups were respectively 79.3%(46/58)and 76.1%(35/46).There was no statistical difference between them(P>0.05)indicating that PPD skin test was not able to diagnose active tuberculosis.In contrast,the positive rate of IFN-γrelease assay for patients reached 95.5%(106/111),while that of healthy controls was 16.3%(46/283).Significant difference existed between them demonstrating that IFN-γrelease assay could detect active tuberculosis with a very high sensitivity(95.5%)and specificity. The overall agreement between PPD skin test and IFN-γrelease assay in detection of latent tuberculous Mycobacterial infection in the controls was 50.0%.However,83.3%of positive people diagnosed by IFN-g release assay were positive for PPD skin test.On the other hand,the overall agreement of these two methods in diagnosis of the active tuberculosis was 72.4%.Among the patients positive diagnosed by IFN-g release assay,77.8%was positive for PPD skin test.Conclusion CFP-10/ESAT-6 specific IFN-γrelease assay is very sensitive and specific in diagnosis of active tuberculosis,while PPD skin test is not able to differentiate BCG vaccination,latent infection and active tuberculosis.

关 键 词：结核 γ干扰素 分枝杆菌 CFP-10 ESAT-6 结核菌素皮内变态反应 PPD 

分 类 号：R521[医药卫生—内科学] R446.6[医药卫生—临床医学]