Expression and purification of Tp0319 recombinant protein of Treponema pallidum and its application in diagnosis of syphilis
- VernacularTitle:梅毒螺旋体Tp0319重组蛋白的表达、纯化及其在临床诊断中的初步应用
- Author:
Yongjian XIAO
;
Ning WU
;
Shuangquan LIU
;
Feijun ZHAO
;
Yimou WU
- Publication Type:Journal Article
- Keywords:
Treponema pallidum;
recombinant protein;
indirect ELISA;
syphilis antibody
- From:
Chinese Journal of Clinical Laboratory Science
1985;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone,express,and purify Tp 0319 outer membrane protein of Treponema pallidum and to develop an indirect ELISA for diagnosing syphilis.Methods The expression plasmid PQE32/Tp 0319 was conventionally constructed.The recombinant Tp 0319 protein was produced in E.coli M15 after induction by IPTG.The Tp 0319 protein was analyzed by SDS-PAGE and Western blotting,and then purified with Ni-NTA affinity chromatography.Indirect ELISA was developed to detect the syphilis antibody in human sera.Results The recombinant plasmid PQE32/Tp 0319 was constructed successfully and the fusion protein with relative molecular weight near 30 000 Dalton was revealed by SDS-PAGE.Western blotting proved that the recombinant protein specifically reacted with anti-Tp antibodies in sera from syphilis patients.The results of the indirect ELISA indicated the sensitivity and the specificity were both 100%.The concordance of 300 sera(150 from blood donors and 150 from syphilis patients)detected in parallel by the ELISA and the TPPA was 95.3%.Conclusions The data suggest that the prepared recombinant protein Tp 0319 of Treponema pallidum has high immunoreactivity.The recombinant protein can be used to develop ELISA kit for diagnosing syphilis.
