OBJECTIVETo 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.

METHODSThe 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.

RESULTSTwo 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).

CONCLUSIONTwo 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.

Adolescent ; Adult ; Chlamydia Infections ; diagnosis ; Chlamydia trachomatis ; pathogenicity ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Male ; Middle Aged ; Urogenital System ; microbiology ; Young Adult

Genital tract infections with ureaplasma urealyticum (UU) and chlamydia trachomatis (CT) are the most frequent sexually-transmitted disease worldwide. UU and CT infections are considered to be the leading cause for infertility and adverse pregnancy outcomes. However, little is known about the specific effect of cervical UU and CT infections on the etiology of female infertility, as well as the pregnancy outcomes of the patients undergoing in vitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET). In order to find the association between cervical UU and/or CT infection and pregnancy outcomes, we conducted a retrospective case-control study on the patients undergoing IVF/ICSI-ET with cervical UU and/or CT infection. A total of 2208 patients who received IVF/ICSI-ET were enrolled in this study. Data on the general conditions, pregnancy history and clinical pregnant outcomes were analyzed in terms of the cervical UU and CT detection. Our results revealed that cervical UU and CT infections were the risk factors for ectopic pregnancy and tubal factor-induced infertility. Moreover, the pregnancy rate, abortion rate, ectopic pregnancy rate and premature birth rate in patients with UU and/or CT infections showed no significant difference when compared with the control group. We recommend that cervical UU and CT detection should be an optional item for infertility patients and clinical UU detection should differentiate the subtypes of cervical UU. Positive cervical UU and CT infections should not be taken as strict contraindications for IVF/ICSI-ET.
Adult ; Chlamydia Infections ; microbiology ; pathology ; Chlamydia trachomatis ; pathogenicity ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Male ; Pregnancy ; Pregnancy Rate ; Premature Birth ; Reproductive Tract Infections ; microbiology ; physiopathology ; Sperm Injections, Intracytoplasmic ; methods ; Ureaplasma Infections ; microbiology ; pathology ; Ureaplasma urealyticum ; pathogenicity

Heat shock proteins (HSP) have been identified as an important factor of a very complex and highly conserved cellular defense mechanism to preserve cell survival under adverse environmental conditions. HSP 60 are immunodominant antigens of microbe such as Chlamydia trachomatis and have a potentiality to become a target antigen due to antigenic similarity between chlamydial and human HSP. This study was conducted to investigate the effects of Vero cell coculture to anti-HSP 60 on the early mouse embryo development in vitro. The 2-cell mouse embryos (ICR) were cultured and mouse embryo development was observed every 24 hr for 3 days. 45% and 22.1% of the embryos cultured in Ham's F-10 plus anti HSP 60 with Vero cells developed to the 4- to 8- cell stage (day 1) and morular stage (day 2) as compared with 29.2% and 2.7% of those cultured without Vero cells respectively. But at day 3, the beneficial effect of Vero cells was not noted. These findings suggest that Vero cells have some roles to overcome the detrimental effect of anti-HSP 60 to some degree. These results suggest that Vero cells coculture will promote reproductive outcome in patient previously sensitized to microbial (e.g. Chlamydia trachomatis) HSP 60.
Vero Cells ; Pregnancy ; Mice, Inbred ICR ; Mice ; Male ; Infertility, Female/etiology/immunology/therapy ; Immunodominant Epitopes ; Female ; Embryonic Development/*immunology ; Coculture Techniques ; Chlamydia trachomatis/immunology/pathogenicity ; Chaperonin 60/*immunology ; Cercopithecus aethiops ; Antigens, Bacterial ; Antibodies, Monoclonal/*administration & dosage ; Animals

PURPOSE: Fitz-Hugh-Curtis Syndrome (FHCS) is a clinical entity characterized by inflammation of the liver capsule associated with genital tract infection. The aim of this study is to provide physicians with clinical suggestions for diagnostic approaches based on a series of patients who were diagnosed with FHCS. MATERIALS AND METHODS: We conducted a retrospective study of patients who were diagnosed with FHCS after presenting to the emergency department (ED). The symptoms, physical examinations, laboratory findings, radiological findings, and progress of the patients were reviewed. RESULTS: During the four-year study period, a total of 82 female patients received a final diagnosis of FHCS in the ED. Chlamydia trachomatis was identified as a pathogen in 89% of the patients. Their clinical characteristics and laboratory findings were described. Fifty-two patients (63.4%) were admitted to the hospital. All of the admitted patients improved after treatment combining antibiotic therapy with conservative care. CONCLUSION: FHCS should be considered as a differential diagnosis for female patients of childbearing age with right upper abdominal pain. Timely diagnosis using biphasic computed tomography (CT) with arterial and portal phases may help ensure adequate medical treatment as well as avoid invasive procedures.
Adolescent ; Adult ; Anti-Bacterial Agents/therapeutic use ; Chlamydia Infections/drug therapy/microbiology/*pathology/radiography ; Chlamydia trachomatis/pathogenicity ; Emergency Service, Hospital ; Female ; Hepatitis/drug therapy/microbiology/*pathology/radiography ; Humans ; Pelvic Inflammatory Disease/drug therapy/microbiology/*pathology/radiography ; Peritonitis/drug therapy/microbiology/*pathology/radiography ; Reproductive Tract Infections/drug therapy/microbiology/pathology/radiography ; Retrospective Studies ; Tomography, X-Ray Computed ; Young Adult