title: Development of ELISAs for the detection of urogenital chlamydia trachomatis infection targeting the pORF5 protein.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  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).
  
  
  CONCLUSIONS
  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.
authors: Department of Microbiology and Immunology, School of Medicine, University of South China, Hengyang 421001, Hunan, China.
authors: Zhong Yu
authors: ZY
authors: Li
authors: Qiu Lin
authors: QL
authors: Huang
authors: Sheng Mei
authors: SM
authors: Su
authors: Guang Ming
authors: GM
authors: Zhong
authors: Yi Mou
authors: YM
authors: Wu
mesh_terms: D000293:Adolescent; D000328:Adult; D002690:Chlamydia Infections / Q000175:diagnosis*; D002692:Chlamydia trachomatis / Q000472:pathogenicity*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D014566:Urogenital System / Q000382:microbiology*; D055815:Young Adult
keywords: Chlamydia trachomatis; DAS-ELISA; Monoclonal antibody; Polyclonal antibody; pORF5