重组CFP-10／ESAT-6抗原ELISA法对活动性肺结核诊断的评估  

The appraisal in diagnosis of active pulmo-tuberculosis of rCFP-10/ESAT-6 fusion protein by ELISA

作　　者：詹枝华[1] 陈颖钰[2] 袁冶[1] 项杰[1] 周金海[1] 郭爱珍[2] 

机构地区：[1]武汉市结核病医院,430083 [2]华中农业大学动物医学院,华中农业大学农业微生物学国家重点实验室,武汉430070

出　　处：《中华微生物学和免疫学杂志》2008年第9期847-850,共4页Chinese Journal of Microbiology and Immunology

基　　金：“973”计划项目（2006CB504401）;武汉市科技攻关课题（20066002056） 志谢：华中农业大学动物医学院博士邓铨涛、匡有吉在本课题基础研究中做了大量的前期工作;武汉市结核病防治所陈军、曾庆志、杜义祥,华中农业大学医院魏武、童庆伟给予了大力支持,在此一并感谢.

摘　　要：目的以rCFP-10／ESAT-6融合蛋白抗原、植物血凝素（PHA）、生理盐水分别处理活动性肺结核患者外周血致敏T淋巴细胞，测定同一标本IFN-γ释放水平，探讨其对结核感染与发病的诊断意义。方法病例组111例为临床确诊肺结核患者，对照组292例为经胸部透视和结核菌素试验（PPD）筛选的大学生。入选对象取血3．0ml，均分为3份，每管1．0ml，分别加入rCFP-10／ESAT-6融合蛋白、PHA和生理盐水，孵育后ELISA法测定IFN-γ的吸光度（A）值及其抗体。结果rCFP-10／ESAT-6融合蛋白处理：IFN-γ测定值病例组x=1．3885±0．6236，对照组x=0．2944±0．0917，组间t′=16．4259，P〈0．05；以0．42为切割值，病例组阳性率93．58％，对照组阳性率13．07％。PHA处理：病例组x=1．2463±0．5541，对照组x=0．5613±0．0640，t′=19．1797，P〈0．05。生理盐水处理：病例组x=0．0772±0．0444，对照组x=0．0290±0．0235，t′=13．9487，P〈0．05。病例组抗体阳性率66．36％，对照组7．19％。结论利用rCFP-10／ESAT-6融合蛋白为特异性抗原，ELISA法测定外周血致敏淋巴细胞IFN-γ释放水平对结核感染诊断的敏感性〉90％，且无论特异性或非特异性处理，活动性肺结核患者IFN-γ释放水平均明显升高，血清抗体阳性率也高于对照组。Objective To sensitize the T-cell in the peripheral blood of the active tuberculosis patients by rCFP-10/ESAT-6 fusion protein, phytohaemagglutinin（PHA） and physiologic saline, and to detect the IFN-γ to approach the significance of the tuberculosis infection. Methods One hundred and eleven patients were diagnosed by clinical definite, 292 undergraduate students were ehosen by X-ray and PPD-selection as volunteers. 3.0 ml of blood was taken from each volunteer, rCFP-10/ESAT-6, PHA and physiologic saline were added into each 1.0 ml, respectively. The A valule and antibody of IFN-γ were assayed by ELISA. Results Treated with rCFP-10/ESAT-6 group ： the A value average of patients group was 1. 3885 ± 0. 6236, students group was 0. 2944 ± 0. 0917. Intergroup t′ = 16. 4259, P 〈 0.05, set 〉 0.42 as cut-off, the positive rate of patients group was 93.58% , students group was 13.07%. Treated with PHA group： the A value average of patients group was 1. 2463. ± 0. 5541, of which the other was 0. 5613 ± 0. 064, t′ = 19. 1797,P 〈 0.05. Treated with physiologic saline group：the A value average of patients group was 0. 0772 -0.0444 ,of which the other was 0. 0290 ±0.0235 ,t′ = 13. 9487, P 〈0.05. All had significant deviation. The antibody positive rate of the patients group was 66.36%, the students group was 7.19%. Conclusion rCFP-10/ESAT-6 as specific antigen, the sensitivity of IFN-γ release assay by ELISA is above 90%. No matter specific or non-specific disposal, the active tuberculosis patients have higher IFN-γrelease level and antibody than the control group.

关 键 词：结核分枝杆菌 结核病 rCFP-10/ESAT-6融合蛋白 酶联免疫吸附法 Γ-干扰素 

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