Development of ELISAs for the detection of urogenital chlamydia trachomatis infection targeting the pORF5 protein / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
;
(12): 169-175, 2013.
Article
in English
| WPRIM
| ID: wpr-320354
ABSTRACT
<p><b>OBJECTIVE</b>To prepare antibodies against pORF5 plasmid protein of Chlamydia trachomatis and develop double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISAs) for the detection of genital C. trachomatis infections.</p><p><b>METHODS</b>The pORF5 protein was expressed in Escherichia coli 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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>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.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Urogenital System
/
Virulence
/
Enzyme-Linked Immunosorbent Assay
/
Chlamydia Infections
/
Chlamydia trachomatis
/
Diagnosis
/
Methods
/
Microbiology
Type of study:
Diagnostic study
Limits:
Adolescent
/
Adult
/
Female
/
Humans
/
Male
Language:
English
Journal:
Biomedical and Environmental Sciences
Year:
2013
Type:
Article
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