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| Expression of recombinant protein of Treponema pallidum and development a sandwich antigen ELISA on serodiagnosis of syphilitics |
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Abstract Objective To clone and express TpN47、TpN17、TpN44.5 and TpN15 out membrane protein of Treponema pallidum, then develop an one-step double antigen sandwich enzyme-linked immunosorbent assay(ELISA) procedure on detecting Tp antibody. Methods The immuno-dominant epotope of TpN47、TpN17、TpN44.5 and TpN15 was amplified by PCR and subcloned into the expression pET-HT-JKM, the recombinant protein was expressed in BL21(DE3)plysS and purified with Ni-NTA affinity chromatography columns. Sandwich antigen ELISA was developed to detect the antibody of Tp in human sera. Results 30 control sera was tested by ELISA. Both the sensitivities and specificities were 100%. While detecting 385 T. pallidum human sera, the sensitivities of ELISA was 98.2% and the specificities was 99.5% compared with the results of the TPPA tests(P＜0.01). The concordance of results between the ELISA test and the TPPA test was 99%. Conclusion The TpN47、TpN17、TpN44.5 and TpN15 out membrane protein of Treponema pallidum were suitable for development of ELISA for serodiagnosis of syphilitics.
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Received: 09 September 2008
Published: 02 July 2009
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Corresponding Authors:
LU Hai-Rong
E-mail: hairongl@szu.edu.cn
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