7种结核分枝杆菌重组蛋白的纯化及血清学特性

    闫冀焕1，史玲莉1，韩伟1，崔鹰1，赵俊伟2，张舒林2

    1.河北国际旅行卫生保健中心，河北 石家庄 050051；2.上海交通大学基础医学院

    摘要：目的分离纯化结核分枝杆菌重组蛋白38-kDa、16-kDa、Mtb81、ESAT-6、CFP10、Rv3425、Rv3391，并比较7种蛋白的免疫学特性，评价其在结核血清学诊断中的应用价值。方法利用间接酶联免疫吸附试验评价确诊结核病人、非结核病人、健康人血清中7种重组蛋白的抗原性。结果成功分离纯化了7种重组蛋白；酶联免疫吸附试验结果表明，7种抗原均能有效区分确诊结核病人、非结核病人和健康人，其中阳性率最高的是ESAT-6为47.3%，特异性最高的是Rv3391为98.3%。联合抗原中阳性率和特异性最高的是CFP10+ ESAT-6 + Rv3391抗原组合。结论 单一抗原中ESAT-6具有较高的免疫原性，联合抗原中CFP10 + ESAT-6 +Rv3391抗原具有很高的阳性率和特异性，在结核诊断具有很高的诊断潜力。

    关键词：结核分枝杆菌；重组蛋白；抗体；抗原；纯化

    中图分类号：R521　文献标识码：B

    The purify and serologic characteristics of 7types of recombinant proteins from mycobacterium tuberculosis

    YAN Ji-huan*, SHI Ling-li, HAN Wei, CUI Ying,ZHAO Jun-wei, ZHANG Shu-lin

    *Hebei International Travel Health CareCenter, Shijiazhuang, Hebei 050000,China

    Abstract:   Objective  To isolate and purify 7 types of proteins(38-kDa、16-kDa、Mtb81、ESAT-6、CFP10、Rv3425、Rv3391) from mycobacteriumtuberculosis, and to compare the immunologic characteristics of 7types of proteins, to evaluate the value of 7 antigens frommycobacterium tuberculosis on the serodiagnosis of tuberculosis(TB).   Methods   7 types of proteins were usedas antigen to screen the sera of tuberculosis (TB) patients(n=203)、healthy controls (n=58)and non-tuberculosis (NTB) patients(n=42) by the enzyme-linked immunosorbent assay(ELISA).   Results   7 types of proteins wereisolated and purified from mycobacterium tuberculosis. There wassignificantly distinction between TB patients, healthy controls andnon-tuberculosis by the 7 proteins. The highest sensitivitiesantibody detected by single antigen was ESAT-6(47.3%), and thehighest specificities was Rv3391(98.3%). The highest sensitivityand highest specificity of antibodies detected by multiple antigenswas CFP10+ESAT-6+Rv3391.   Conclusion   As asingle antigen of proteins from mycobacterium tuberculosis, theimmunogenicity of ESAT-6 is better than the other 6 antigens. Themultiple antigens of CFP10+ ESAT-6+ Rv3391 can increase thesensitivity and specificity of TB antibody detection, and it may bevaluable in the diagnosis of TB.

    Key words:   Mycobacteriumtuberculosis; Recombinant proteins; Antigens; Antibody; Purify

    《中国国境卫生检疫杂志》2013年6月刊