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

发布时间：2018-04-15 10:25

本文选题：结核分枝杆菌 + 抗体　；参考：《延边大学》2009年硕士论文

【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。为开发新型结核分枝杆菌诊断性试剂,提供理论依据。 [方法]应用硫酸铵盐析作用、疏水作用和阴离子交换作用从结核分枝杆菌培养滤过液中分离纯化30-kDa和38-kDa蛋白:应用金属螯合NI~(2+)-NTA琼脂亲和层析作用从结核分枝杆菌培养滤过液中分离纯化ESAT-6、Rv3872和HBHA蛋白。在肺结核患者和正常对照组血清中,利用间接酶联免疫吸附试验,评价五种结核分枝杆菌分泌性蛋白及其三种联合蛋白的免疫原性。 [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。
[Abstract]:Separation and purification of five secreted proteins of Mycobacterium tuberculosis to (30-kDa, 38-kDa, ESAT-6, Rv3872, and, HBHA) between the five proteins and three combined protein (38-kDa-30-kDa Ag, 38-kDa-ESAT-6 Ag, 38-kDa-HBHA Ag) on the immunological characteristics, and to evaluate its application value in serological diagnosis of tuberculosis. For the development of new Mycobacterium tuberculosis diagnostic reagents, and provide a theoretical basis.
[method] the application of ammonium sulfate precipitation, hydrophobic interaction and anion exchange separation and purification of 30-kDa and 38-kDa culture filtrate protein from Mycobacterium tuberculosis: application of metal chelating NI~ (2+) -NTA agar affinity chromatography purification effect of ESAT-6 filtration liquid from Mycobacterium tuberculosis culture, Rv3872 and HBHA protein in patients with pulmonary tuberculosis. And the normal control group in serum by indirect enzyme linked immunosorbent assay, the immunogenicity of secretory protein and its three joint evaluation of five protein of Mycobacterium tuberculosis.
[results] the successful purification of 5 kinds of secretory proteins. And in ELISA, 38-kDa antigen and other four secreted proteins from Mycobacterium tuberculosis, and has high sensitivity and specificity, especially in the induction of immune response to the strongest anti IgM antibody (P < 0.0001), its sensitivity and specificity of 44.44% and 95%. respectively in 38-kDa and 30-kDa and the sensitivity of combined antigen in up to 80%, and also has a high specificity. The results also showed that ESAT-6 antigen and heparin binding hemagglutinin adhesin antigen (HBHA), and in three kinds of antibody response, with similar sensitivity and specificity of.Rv3872 antigen than the other four antigens of sensitivity and specificity are low.
[conclusion]1. as a single antigen of Mycobacterium tuberculosis secretory protein, the immunogenicity of 38-kDa antigen is superior to the other four antigens (30-kDa, ESAT-6, Rv3872 and HBHA),.2.38-kDa and 30-kDa combined antigen. It is hoped to become an effective candidate antigen for new diagnostic reagent of tuberculosis serology.

【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392

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