五种结核分枝杆菌分泌性蛋白的纯化及其血清学特性的研究

【作者】 李金花； 林星； 陈爽； 张庆镐；

【机构】 延边大学基础医学院免疫学与病原生物学教研部； 延边大学附属医院胸外科；

【摘要】 目的分离纯化五种结核分枝杆菌分泌性蛋白(30-kDa,38-kDa,ESAT-6,Rv3872 and HBHA),比较这五种蛋白及三种联合蛋白(38-kDa-30-kDa Ag,38-kDa-ESAT-6 Ag,38-kDa-HBHA Ag)的免疫学特性,并评价其在结核病血清学诊断方面的应用价值。方法在肺结核病患者和正常对照组血清中,利用间接酶联免疫吸附试验,评价五种蛋白及三种联合蛋白的抗原性。结果成功分离纯化了5种分泌性蛋白。并且在酶联免疫吸附试验中,38-kDa抗原与其他四种结核分枝杆菌分泌性蛋白比较,具有较强的敏感性和特异性,尤其在抗IgM抗体的诱导免疫反应最强(p<0.0001),其敏感性和特异性分别为44. 44%和95%。而在38-kDa与30-kDa联合抗原中敏感性高达80%,并且也具有较高的特异性。结果还显示出,ESAT-6抗原和肝素结合血凝黏附素(HBHA)抗原,在与三种抗体的反应中,具有相似的敏感性与特异性。Rv3872抗原比其他四种抗原的敏感性和特异性均低。结论1.作为结核分枝杆菌分泌性蛋白单一抗原,38-kDa抗原的免疫原性优于其他四种抗原(30-kDa, ESAT-6,Rv3872 and HBHA)。2.38-kDa与30-kDa联合抗原,有希望成为结核病血清学新型诊断试剂有效的候选抗原。更多还原

【Abstract】 Objective To purify the five secreted antigens(30-kDa,38-kDa,ESAT-6,Rv3872 and HBHA) from M.tuberculosis, and compare the humoral immune responses induced by the five secreted proteins and the combination of the individual antigens (38-kDa plus 30-kDa Ag,38-kDa plus ESAT-6 Ag,38-kDa plus HBHA Ag).Methods The antibody(IgG,IgM,IgA) responses against five Mycobacterium tuberculosis antigens and the combination proteins are estimated in sera from TB patients and healthy controls, using the indirect enzyme-linked immunoabsorbant assay(ELISA).Results In ELISA test,the 38-kDa antigen has stronger sensitivity and specificity than the other four secreted proteins,especially the response with IgM antibody(p<0.0001 ).The sensitivity and specificity of 38-kDa antigen are 44.44%and 95%,respectively.However,the sensitivity increased to 80%when the combination of 38-kDa and 30-kDa antigen is measured.Specificity remained high level when the combination of individual antigens is used. The sensitivity and specificity of ESAT-6 Ag is similar to heparin-binding hemagglutinin adhesion(HBHA) antigen with the responses of three anti-human antibodies.The Rv3872 antigen has lower sensitivity and specificity than the other four secreted proteins.Conclusions As a single M.tuberculosis secreted protein,the immunogenicity of 38-kDa antigen is better than the other four(30-kDa, Rv3872,ESAT-6,HBHA) antigens in the serological test.The 38-kDa Ag,in combination with 30-kDa Ag,is promising candidates for immunodiagnostic assays specific for tuberculosis.更多还原