CAJ | 학술논문

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.
목적 구건결핵분지간균CFP10、MPT48화TB8.4융합단백,평개기용우결핵병혈청학검측적개치.방법 선취2008년상해시폐과의원폐결핵환자150례,비결핵폐부질병환자70례,건강체검자103명,채집혈청표본공323빈.구건결핵분지간균CFP10、MPT48화TB8.4융합표체재체.융합단백경면역인적분석후,채용매련면역흡부시험검측혈청표본,운용Medcale11.5대결과진행통계학분석.결과 성공구건중조질립pET21a-cfp10-mpt48-tb8.4,획득95％이상순도적융합단백.면역인적법발현융합단백여혈청항체발생면역반응.융합단백검측혈청항체민감도56.7％(85/150),특이도90.8％(157/173),기중비결핵병환자특이도위85.7％ (60/70),건강체검자특이도위94.2％(97/103).결론 CFP10-MPT48-TB8.4융합단백소구유적면역반응성,유가능성위결핵병혈청학진단적후선항원.
Objective 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.