摘要目的 构建结核分枝杆菌CFP10、MPT48和TB8.4融合蛋白,评价其用于结核病血清学检测的价值.方法 选取2008年上海市肺科医院肺结核患者150例,非结核肺部疾病患者70例,健康体检者103名,采集血清标本共323份.构建结核分枝杆菌CFP10、MPT48和TB8.4融合表达载体.融合蛋白经免疫印迹分析后,采用酶联免疫吸附试验检测血清标本,运用Medcale11.5对结果进行统计学分析.结果 成功构建重组质粒pET21a-cfp10-mpt48-tb8.4,获得95％以上纯度的融合蛋白.免疫印迹法发现融合蛋白与血清抗体发生免疫反应.融合蛋白检测血清抗体敏感度56.7％(85/150),特异度90.8％(157/173),其中非结核病患者特异度为85.7％ (60/70),健康体检者特异度为94.2％(97/103).结论 CFP10-MPT48-TB8.4融合蛋白所具有的免疫反应性,有可能成为结核病血清学诊断的候选抗原.

abstractsObjective To construct the recombinant plasmid of protein CFP10-MPT48-TB8.4 of Mycobacterium tuberculosis and to investigate the diagnosis potential of this fusion protein in tuberculosis serodiagnosis.Methods The recombinant fusion protein CFP10-MPT48-TB8.4 was expressed, and identified by Western blot.The ELSIA based on the purified fusion protein was done,and used for screening in 230 cases of clinical serum samples including pulmonary tuberculosis patients ( n =150 ),pulmonary disease patients other than tuberculosis (n =70) and health controls (n =103 ).The test result was analyzed by Medcale11.5 software.Results The fusion protein CFP10-MPT48-TB8.4 was successfully expressed with a purity over 95％.Specific immunogenicity of the recombinant protein was confirmed by Western blot.The overall sensitivity and specificity obtained of ELISA were 56.7％ (85/150) and 90.8％ ( 157/173 ),respectively.The specificity was 85.7 ％ (60/70) in non-tuberculosis group and 94.2％ (97/103 ) in healthy group,respectively.Conclusion The recombinant protein of CFP10-MPT48-TB8.4 has a high sensitivity and specificity and may be a potential candidate antigen in tuberculosis serodiagnosis.