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Detection of bacterial vaginosis, Trichomonas vaginalis infection, and vaginal Candida infection: a comparative study of methods of extracting exudates, with and without a speculum, during pregnancy.

PURPOSE: To compare the collection of vaginal exudates with and without the use of a speculum to diagnose vaginal infections. MATERIAL AND METHODS: We examined 45 patients with vaginal discharge. Two vaginal swabs were taken, one with and the other without a speculum. Both were examined by wet mount microscopy to detect bacterial vaginosis (BV), Trichomonas vaginalis, and the presence of blastospores with or without pseudomycelium. RESULTS: A total of 90.9% of patients whose vaginal exudates were negative for BV when extracted with a speculum were also negative in the swab taken without a speculum. In the case of BV-positive exudates using a speculum, the correspondence was 90.5% with the swabs taken without a speculum. Among those exudates obtained with a speculum that were negative for Candida, 92.6% of those obtained without a speculum tested negative. In patients with pseudomycelium, when a speculum was used, 100% also presented a diagnosis of pseudomycelium from the swabs taken without a speculum. A concordance test between the techniques involving the use or nonuse of a speculum was performed for each of the diagnoses showed a good agreement according to the observed Kappa statistics (0.8467 and 0.8396 for BV and Candida, respectively). CONCLUSIONS: A very good agreement between the results obtained with and without the use of a speculum was observed. Swabbing without the use of a speculum is especially convenient for pregnant women who require frequent testing for these types of infections during their pregnancies.

Candidiasis, Vulvovaginal↗

Improved diagnosis of Trichomonas vaginalis infection by PCR using vaginal swabs and urine specimens compared to diagnosis by wet mount microscopy, culture, and fluorescent staining.

Four vaginal cotton swab specimens were obtained from each of 804 women visiting the outpatient sexually transmitted disease clinic of the Erasmus University Medical Center Rotterdam, Rotterdam, The Netherlands, for validation of various forms of Trichomonas vaginalis diagnostic procedures. One swab specimen was immediately examined by wet mount microscopy, a second swab was placed in Kupferberg's Trichosel medium for cultivation, and two swabs were placed in phosphate-buffered saline (PBS), pH 7.2. The resulting PBS suspension was used for direct staining with acridine orange and fluorescence microscopy, inoculation of modified Diamond's culture medium, and a PCR specific for T. vaginalis. A total of 70 samples positive in one or more of the tests were identified: 31 (3.8%) infections were detected by wet mount microscopy, and 36 (4.4%) were identified by acridine orange staining, as opposed to 40 (4.9%) and 46 (5.7%) positives in modified Diamond's and Trichosel media, respectively. PCR was positive for 61 (7.5%) samples. Secondly, from each of 200 women were obtained a urine sample and a vaginal cotton swab specimen, and 200 urine samples were obtained from men. For the women, 15 (7.4%) of the samples showed a positive result for either the wet mount (n = 1), Trichosel culture (n = 6), PCR on the vaginal swab sample (n = 10), or PCR on the urine specimen (n = 11). Four men (2%) were diagnosed with a T. vaginalis infection. Thus, PCR appears to be the method of choice for the detection of genital infections with T. vaginalis.

Animals↗

The vagina of women infected with Trichomonas vaginalis has numerous proteinases and antibody to trichomonad proteinases.

BACKGROUND: Patients with trichomoniasis have serum antibody to numerous T. vaginalis cysteine proteinases, indicating that the proteinases are expressed in vivo. It was important, therefore, to examine for the presence of soluble trichomonad proteinases and/or antibody to the proteinases in the vagina of infected women. METHODS: Vaginal washes (VWs) from 20 women were examined for the presence of proteinases by electrophoresis using acrylamide co-polymerised with gelatin as the indicator system. Antibody to proteinases in VWs was detected by an immunoprecipitation assay involving protein A-bearing Staphylococcus aureus first coated with anti-human immunoglobulin G (IgG) antibody, which was then added to VWs. For VWs having soluble proteinases, the bacteria were used to determine whether immune complexes between antibody and proteinases were present. VWs without soluble proteinases were incubated with the anti-human IgG treated bacteria before adding to detergent extracts of T. vaginalis. Individual isolates from the patients examined in this study were also analysed by one- and two-dimensional electrophoresis for their proteinase content. Finally, VWs were from patients without any history of other sexually transmitted diseases (STDs) as well as from individuals having numerous other STDs, including yeast, group B streptococcus, chlamydia, and syphilis. RESULTS: Approximately one-third of patients had soluble proteinases in the VWs; the remaining two-thirds (70%) of patients and normal women had no detectable proteinases in VWs. Half of the patients without soluble proteinases had IgG which, when bound to S. aureus, immunoprecipitated many proteinases from a detergent extract of T. vaginalis. All soluble proteinases and those precipitated from trichomonal extracts were inhibited by inhibitors of cysteine proteinases. Finally, patients having trichomoniasis in addition to numerous other STD agents, including yeast, group B streptococcus, chlamydia, and syphilis did not have soluble proteinases in VWs. Equally noteworthy, some patients with soluble proteinases in VWs did not have other detectable STD agents. CONCLUSIONS: Proteinases were detected in the vagina of some patients with trichomoniasis, and in most cases the proteinases were complexed with IgG, which was precipitated by S. aureus. Patients without soluble proteinases in VWs also had antibody specifically to trichomonad proteinases, again demonstrating both the expression and immunogenic nature of the proteinases in vivo. The absence of soluble proteinases in normal women and in patients having other STD agents as well as the presence of proteinases in VWs of patients without other detectable STD pathogens reinforced the idea that the proteinases were of T. vaginalis parasite origin. The findings of this study indicate that proteinases may be important to the T. vaginalis-host interrelationship.

Animals↗

Trichomonas vaginalis infections in Sydney: laboratory diagnosis and prevalence.

The prevalence of trichomoniasis in Sydney was determined by examining 1070 sexually active females in three different practices. Infection rates ranged from 17.8% in a sympatomatic venereal-disease clinic group to 0.5% in non-promiscuous family-planning clinic patients. In each of these categories, the infestation rate, when compared with overseas reports, was lower but no explanation for this difference is apparent. Of the various laboratory methods used to detect Trichomonas vaginalis, the most sensitive was a culture method. The most convenient and reliable microscopy technique was phase-contrast microscopy which detected 80% of those with positive culture results. Stuart's Transport Medium will maintain trichomonad's viability for at least 24 hours and allow vaginal specimens to be reliably transported for examination.

Australia↗

Antibody in sera of patients infected with Trichomonas vaginalis is to trichomonad proteinases.

BACKGROUND: A recent report demonstrated the immunogenic character of the cysteine proteinases of Trichomonas vaginalis. It was of interest, therefore, to examine for the presence of serum anti-proteinase antibody among patients with trichomoniasis. METHODS: An immunoprecipitation assay was used involving protein A-bearing Staphylococcus aureus first coated with the IgG fraction of goat anti-human Ig and then mixed with individual sera of patients to bind human antibody. These antibody-coated bacteria were then added to detergent extracts of T vaginalis. Bound immune complexes on S aureus were washed and solubilised for electrophoretic analysis on acrylamide copolymerised with gelatin for detection of proteinase activity. RESULTS: Sera from patients (50/50), but none from sera of normal, uninfected women, possessed IgG to numerous trichomonad cysteine proteinases. The presence of this serum anti-proteinase antibody disappeared after drug treatment and cure of patients of the T vaginalis infection. CONCLUSIONS: The commonality of the anti-proteinase antibody in the sera of patients with trichomoniasis provided evidence for the expression of the same repertoire of parasite proteinases during infection. These observations have important implications for the in vivo relevance of the proteinases and indicate that strategies to use a specific serum antibody response for diagnosis of this infection may be possible.

Animals↗

Diagnosis of Trichomonas vaginalis in adolescent females: InPouch TV culture versus wet-mount microscopy.

PURPOSE: This study compared the InPouch TV culture to wet-mount, Diamond's culture medium, and Papanicolaou (Pap) smear for the diagnosis of trichomonas infection in sexually active adolescents. METHODS: A total of 467 subjects were recruited among 12-18-year-old girls who received pelvic examinations at two urban adolescent clinics. All girls were tested by wet-mount and InPouch TV. In addition 339 of 467 had cultures in Diamond's medium and 366 of 467 had Pap smears. Specimens were collected for InPouch TV and Diamond's cultures and read at 24-48 h and 5 days, and in the case of Diamond's cultures, also at 7 days. In a subset of subjects (268 of 467) who had all four tests done, sensitivities and specificities were calculated using Diamond's culture as the "gold standard." RESULTS: In the 467 subjects, 73 (15.6%) tested positive for trichomonas by at least one method. In the subset with all four tests done, sensitivities of the wet-mount and InPouch TV were 36% and 81%, respectively; while that of the Pap smear was 56%. The culture media were equally efficient in identifying Trichomonas vaginalis. There were no differences found between subjects with or without trichomonas infections in gynecological symptoms, previous history of sexually transmitted diseases, or use of a condom at last intercourse. CONCLUSIONS: InPouch TV culture is a good diagnostic method for T. vaginalis because of its long shelf-life, relatively low expense, and high sensitivity (over twice as sensitive as wet-mount).

Adolescent↗