Objective To prepare antibodies against pORF5 plasmid protein of Chlamydia trachomatis develop double-antibody sandwich enzyme-linked immunosorbent assays （DAS-ELISAs） for detection of genital C. trachomatis infecti...Objective To prepare antibodies against pORF5 plasmid protein of Chlamydia trachomatis develop double-antibody sandwich enzyme-linked immunosorbent assays （DAS-ELISAs） for detection of genital C. trachomatis infections. and the Methods The pORF5 protein was expressed in Escherichia coil and used to immunize BALB/c mice and New Zealand rabbits to produce monoclonal antibodies （mAbs） and polyclonal antibody （pAb） for DAS-ELISAs. Clinical samples from 186 urogenital infection patients （groups I） and 62 healthy donors （groups II） were detected in parallel by the DAS-ELISAs developed in this study and by IDEIA PCE commercial ELISA. Results Two hybridoma cell lines, named 2H4 and 4E6, stably secreting specific mAbs against pORF5 were obtained. The mAb 2H4 was recognized by 32 （17.20%, positive recognition rate） and 25 （13.44%）, mAb 2H4 by 0 （0%） and 2 （3.22%） samples from groups I and II, respectively. The sensitivities of mAbs 2H4 and 4E6 were 92.11% and 77.78% and the specificities were 100% and 96.88%, respectively in relation to the IDEIA PCE commercial ELISA. The sensitivities of detection for the DAS-ELISAs were 10 ng/mL （based on 2H4） and 18 ng/mL （based on 4E6）. Conclusion Two DAS-ELISAs were developed in this study that provided a feasible and effective assay that could be considered alternative tools for the serodiagnosis of C. trachomatis infection.展开更多
基金supported by grants from the National Natural Science Foundation of China (30970165 and 81102230)Hunan Provincial Natural Science Foundation of China (09JJ3059)Team Project for the Technology Innovation of Higher Education of Hunan province, China, 2010
文摘Objective To prepare antibodies against pORF5 plasmid protein of Chlamydia trachomatis develop double-antibody sandwich enzyme-linked immunosorbent assays （DAS-ELISAs） for detection of genital C. trachomatis infections. and the Methods The pORF5 protein was expressed in Escherichia coil and used to immunize BALB/c mice and New Zealand rabbits to produce monoclonal antibodies （mAbs） and polyclonal antibody （pAb） for DAS-ELISAs. Clinical samples from 186 urogenital infection patients （groups I） and 62 healthy donors （groups II） were detected in parallel by the DAS-ELISAs developed in this study and by IDEIA PCE commercial ELISA. Results Two hybridoma cell lines, named 2H4 and 4E6, stably secreting specific mAbs against pORF5 were obtained. The mAb 2H4 was recognized by 32 （17.20%, positive recognition rate） and 25 （13.44%）, mAb 2H4 by 0 （0%） and 2 （3.22%） samples from groups I and II, respectively. The sensitivities of mAbs 2H4 and 4E6 were 92.11% and 77.78% and the specificities were 100% and 96.88%, respectively in relation to the IDEIA PCE commercial ELISA. The sensitivities of detection for the DAS-ELISAs were 10 ng/mL （based on 2H4） and 18 ng/mL （based on 4E6）. Conclusion Two DAS-ELISAs were developed in this study that provided a feasible and effective assay that could be considered alternative tools for the serodiagnosis of C. trachomatis infection.
