2. Materials and Methods

The population studied consisted of asymptomatic nonpregnant women of reproductive age presenting to the Obstetrics and Gynaecology Outpatient Clinic of Aretaieio University Hospital for a routine Pap smear test between January 2008 and May 2012 and who gave consent to be included in this study. The Ethics Committee of Aretaieio Hospital approved the study, and informed consent forms were obtained from all subjects.

Women were counselled to present to the hospital 10 to 20 days after the first day of their menstrual cycle without using vaginal douche or medication for at least a month. The smears showing inflammatory changes or no inflammation were included in this study, while inadequate smears and smears showing any atypical morphology were excluded. Inflammatory changes as detected on a Pap stained smear included the presence of cells with enlarged nuclei, pyknosis or karyorrhexis, perinuclear halos, and vacuoles. Often increased numbers of polymorphonuclear leukocytes or neutrophils and parabasal cells with generalised eosinophilia of the cells were recorded. Furthermore, the presence of epithelial cells covered with blue-stained coccoid bacteria on the stained cervical smear together with a decreased number or a lack of lactobacilli represented findings suggestive of bacterial vaginosis.

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Asymptomatic women with or without inflammatory changes on their Pap smear were recalled for cultures. Genital tract samples (vaginal and cervical) were available for analysis. A wet mount as well as a gram-stained smear was examined under microscope to screen for Trichomonas vaginalis and to obtain valuable information about the microorganisms (yeasts and bacteria) present as well as for the diagnosis of bacterial vaginosis using both Amsel and Nugent criteria [8, 9]. The women were tested for the presence of different aerobic bacteria, that is, Candida species, group B streptococcus, Gardnerella vaginalis, and Neisseria gonorrhoeae, and anaerobic bacteria. Thus, clinical specimens collected from patients were inoculated onto appropriate plates for standard aerobic and anaerobic cultures and incubated at 37°C for 24 h and 48 h, respectively. The isolated pathogens were identified using the automated system VITEK 2 (BioMerieux, Marcy l’Etoile, France). Furthermore, the presence of Chlamydia trachomatis as well as Ureaplasma urealyticum and Mycoplasma hominis, in the specimens studied, was determined using the COBAS AMPLICOR Chlamydia trachomatis test (Roche Diagnostics, USA) and Mycoplasma IST2 (BioMerieux, Marcy l’Etoile, France), respectively.

Statistical analysis was performed using chi square test, and values ≤0.05 were considered as significant.

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