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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。
Previous studies have established the IFN-γ release response of peripheral blood mononuclear cells based on the Mycobacterium tuberculosis specific antigen CFP-10 / ESAT-6 fusion protein. The purpose of this study was to confirm the potential of the method in active tuberculosis and TB latent infection The significance of diagnosis. Participants included 111 cases of active tuberculosis (case group) and 283 freshmen (healthy controls) admitted to a local hospital. Heparin anticoagulant blood was collected and added to the cell culture wells containing Mycobacterium tuberculosis antigen CFP-10 / ESAT-6, phytohemagglutinin and non-irritant (PBS), cultured overnight, and the serum was collected the next day for IFN- . Meanwhile, 58 patients and 46 healthy control volunteers were given tuberculin (PPD) intradermal allergy. The positive rates of intradermal PPD in case group and healthy control group were 79.3% (46/58) and 76.1% (35/46) respectively, with no significant difference (P> 0.05), indicating that PPD intradermal allergy can not be used in Detection of active tuberculosis. In the case group, the positive rate of IFN-γ released in vitro was 95.5% (106/111), while the positive rate of healthy control group was 16.3% (46/283). There was significant difference between the two groups, indicating that in vitro release of IFN-γ Response to diagnosis of active tuberculosis with high sensitivity (95.5%) and good specificity. In the healthy group, the total coincidence rate of IFN-γ release in vitro and PPD intradermal allergy was 50.0%, and 83.3% of IFN-γ release-positive in vitro was PPD intradermal allergy; in the case group , The total coincidence rate between the two was 72.4%, 77.8% of IFN-g positive in vitro release-positive PPD intradermal allergy. CFP-10 / ESAT-6 specific IFN-γ release in vitro diagnosis of active tuberculosis has a high sensitivity and specificity, PPD intradermal allergy due to BCG immunity and other factors in our country can not be used to diagnose active tuberculosis Mycobacterium tuberculosis infection.