重组梅毒螺旋体多表位抗原的改进及应用 (pdf)

    【摘要】 目的 对梅毒螺旋体抗原表位进一步优化，以满足梅毒检测试剂的需要。 方法 用信息生物学软件对梅毒螺旋体TpN47抗原表位进行分析，在我们以前研究工作的基础上，延伸其长度，用化学合成法获得目的基因，插入到pBVIL1载体中，构建pBVIL1/TpN47表达质粒。将新获得的TpN47基因与本研究室保存的TpN15、TpN17、TpN44.5基因片段相连接，进行嵌合表达，经纯化后用双抗原夹心法测其活性。 结果 经改进的重组梅毒螺旋体多表位嵌合抗原，用于检测国家新一代的参考品，其阳性及阴性符合率均为100%，4份灵敏度符合参考品的要求。 结论 所构建的pBVIL1/TpN15+TpN44.5+TpN17+TpN47嵌合质粒，在E.coli中获得了高效表达，经测定，此抗原可满足新一代梅毒检测试剂的需要。

    【关键词】 梅毒螺旋体；TpN47；嵌合抗原；参考品

    Improvement based on recombinant multi-epitope chimeric antigen Treponema pallidum and its application

    SONG Xiao-gou，WANG guo-hua，CHEN Kun，et al.Institute of Basic Medical Sciences, AMMS, Beijing 100850,China

    【Abstract】 Objective In order to detect Kit antibodies of syphilis，the epitope chimeric antigen of Treponema pallidum were optimized further. Methods Treponema pallidum TpN47 antigen epitope were analyzed by information biology software,to extend its length, then linked with TpN15, TpN17, TpN44.5 antigens gene fragment. The recombinant Treponema pallidum multi-epitope chimeric antigens was expressed in E.coli. The Activity of multi-epitope chimeric antigens were detected by sandwich antigen enzyme-linked immunosorbent assay (ELISA). Results Improvement based on recombinant multi-epitope chimeric antigen Treponema pallidum antigens were used newly panel of antibodies syphilis, results of detection were both positive and negative coincidence ratio of 100%, 4 serial sensitivity consistent with panel. Conclusion Recombinant of multi-epitope chimeric antigen of Treponema pallidum was highly expressed in E.coli. Satisfy requirememnt for new panel of syphanaly antibodies. ......