【摘要】 目的表达梅毒螺旋体(Tp)重组蛋白Tp0971(rTp0971),评价其作为候选抗原在梅毒血清学诊断中的意义。方法 PCR扩增Tp0971基因,构建原核重组体pET-28a/Tp0971并诱导表达蛋白,Western blot检测表达产物的抗原性;建立rTp0971-ELISA,检测238份TPPA阳性和120份TPPA阴性血清的Tp特异性IgG,并与TPPA法比较。结果成功高效表达了rTp971;Western blot显示纯化的rTp0971能被梅毒患者血清识别。用rTp0971-ELISA检测临床血清标本的特异性为100%,敏感性为89.1%,总符合率为92.7%。结论 rTp0971有较好的抗原性,可望作为梅毒血清学检测的候选诊断抗原。更多还原

【Abstract】 Objective To express recombinant protein Tp0971(rTp0971) of Treponema pallidum(Tp)and to evaluate its significance in syphilis serodiagnosis as a potential diagnostic antigen.Methods The Tp0971 gene was amplified by PCR.The prokaryotic recombinant vector pET-28a/Tp0971 was constructed and expression of rTp0971 was induced.Antigenicity of purified rTp0971 expressed in E.coli Rosetta was analyzed with Western blot.Indirect rTp0971-ELISA was developed for detection of specific anti-Tp IgG in 238 TPPA-positive clinical serum samples and 120 TPPA-negative serum samples.rTp0971 was evaluated in comparison to TPPA as a potential diagnostic antigen for syphilis serodiagnosis.Results rTp971 protein was efficiently expressed in E.coli.Western blot revealed that purified rTp971 was present in the sera of syphilis patients.When detecting rTp971 in clinical serum samples,rTp0971-ELISA had a specificity of 100%,sensitivity of 89.1%,and rate of concordance of 92.7% when compared to TPPA.Conclusion rTp0971 has good antigenicity and offers promise as a potential diagnostic antigen for syphilis serologic tests.更多还原