酶联免疫斑点试验检测技术在血液病合并痰涂片抗酸杆菌阴性结核病诊断中的应用  

Implication of ELISPOT assay for diagnosis of tuberculosis in sputum-negative patients with hematologic diseases

作　　者：叶丽萍[1] 梁艳 施兵[1] 刘丽辉[1] 金建刚[1] 张玉珠[1] 胡文清[1] 刘明娟[1] 阳幼荣 陈虎[3] 吴雪琼 

机构地区：[1]解放军总医院第二附属医院血液科,北京100091 [2]全军结核病研究所 [3]军事医学科学院附属医院造血干细胞移植中心

出　　处：《中国感染与化疗杂志》2010年第1期44-48,共5页Chinese Journal of Infection and Chemotherapy

摘　　要：目的评价酶联免疫斑点试验(ELISPOT)检测外周血中结核分枝杆菌抗原特异性γ干扰素水平,在诊断免疫功能低下的血液病合并痰涂片抗酸杆菌阴性的结核病患者时的应用价值。方法采用以结核分枝杆菌特异性重组ESAT-6和CFP-10融合蛋白为刺激抗原的ELISPOT检测技术,分析了51例发热的血液病怀疑合并痰涂片抗酸杆菌阴性结核病患者的外周血,作能分泌结核特异的γ干扰素的T淋巴细胞测定,以传统的诊断方法为对照。结果确诊血液病合并结核病24例,排除活动性结核病27例;ELISPOT的灵敏度为73.6%,高于PPD皮肤试验的54.5%(P<0.05),高于抗结核抗体试验的37.5%(P<0.01);准确度82.6%高于抗结核抗体的62.5%(P<0.05),高于PPD的77.77%;特异度88.88%,与抗结核抗体检测(87.5%)及PPD(93.75%)相似;ELISPOT与抗结核抗体、PPD试验结果联合分析,其诊断率为79.2%。此外,ELISPOT检测结果与性别、年龄、结核病史、抗结核抗体、机体免疫状态、外周血淋巴细胞数等影响因素之间的相关性进行了初步探讨。结论ELISPOT是血液病患者合并活动性结核的重要辅助诊断工具。Objective To evaluate the utility of ELISPOT assay in the diagnosis of tuberculosis,especially in sputum-negative（TB）,febrile patients with hematologic diseases.Methods ELISPOT assay used Mycobacterium tuberculosis-specific fusion protein of recombinant ESAT-6 and CFP-10 as the stimulating antigen.This ELISPOT assay was employed to test 51 febrile sputum-negative patients with hematologic diseases for suspicious tuberculosis.T lymphocytes capable of secreting TB-specific gamma-interferon were analyzed in the peripheral blood of these patients.The conventional diagnostic tools（3 anti-TB antibodies and PPD） were used as control.Results Hematologic diseases complicated with tuberculosis were confirmed in 24 of the 51 febrile patients.TB was excluded in the remaining 27 patients.ELISPOT showed higher sensitivity（73.6%） than PPD skin test（54.5%,P0.05） and the 3 anti-TB antibodies（37.5%,P0.01）.The accuracy of ELISPOT was the highest（82.6%）,followed by PPD（77.77%） and 3 anti-TB antibodies（62.5%,P0.05）.The specificity of ELISPOT lied between that of PPD（93.75%） and anti-TB antibodies（87.5%）.ELISPOT assay in combination with anti-TB antibodies and PPD raised the positive rate to 79.2%.In addition,the authors discussed the results in relation to such factors as sex,age,TB history,anti-TB antibodies,immune status and count of peripheral lymphocytes.Conclusions ELISPOT assay may serve as an important supplementary tool in the diagnosis of tuberculosis in patients with hematologic diseases.

关 键 词：血液病 结核病 诊断 酶联免疫斑点试验 

分 类 号：R55[医药卫生—血液循环系统疾病] R446.6[医药卫生—内科学] R52[医药卫生—临床医学]