Cell culture compared with broth for detection of Trichomonas vaginalis

Abstract

Trichomonas vaginalis can be grown in cell culture. We studied the growth kinetics of T. vaginalis in McCoy cell culture compared with that in a conventional broth medium (Diamond TYI-S-33 medium supplemented with 10% heat-inactivated bovine serum [TYI]). In the presence of McCoy cells and two parts cell culture medium to one part TYI, a peak concentration of 2 x 106 to 6 x 106 T. vaginalis per ml was consistently achieved with inocula as low as three T. vaginalis cells per ml. Without cells, this medium did not support growth of T. vaginalis. T. vaginalis in TYI in 1-ml vials with or without McCoy cells demonstrated poor growth. In tubes containing 10 ml of TYI, inocula grew to 2 x 106 to 6 x 106 T. vaginalis per ml, but at least 3 x 101 T. vaginalis per tube was required to initiate growth. Thus, in vitro, cell culture was more sensitive than TYI broth in detecting low numbers of T. vaginalis. In a subsequent clinical comparison of broth and cell culture for isolation of T. vaginalis from 188 vaginal specimens and 21 urethral specimens from men, the results were in agreement for 206 specimens (98.6%). There were no situations in which culture was negative and a saline preparation showed motile trichomonads. For women, using a positive culture as the indicator of true positivity, the sensitivity of detection of T. vaginalis was 83% with the Pappenheim stain and 77% with saline preparations. In tests with a limited number of men, the Pappenheim stain was neither sensitive nor specific for detection of T. vaginalis in comparison with cultures or saline preparations. These studies show that cell culture can be used for isolation of T. vaginalis from clinical specimens; it gave results comparable to those of broth culture for the group of mainly symptomatic womèn. Further studies should be performed to determine its utility in clinical populations such as asymptomatic women and men with and without symptoms, in which T. vaginalis is more likely to be present in low nuffibers.

Inducible immunity to Trichomonas vaginalis in a mouse model of vaginal infection

“…T. vaginalis isolates obtained from vaginal secretions of women with acute vaginitis were grown as previously described (19) in 10 ml of Diamond's TYI-S-33 medium (TYI), pH 6.2 (14), supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Gibco Laboratories, Life Technologies Inc., Grand Island, N.Y.), 100 U of penicillin per ml, 100 g of streptomycin per ml (penicillin-streptomycin solution, Gibco Laboratories), and 2.5 g of amphotericin B (Gibco Laboratories) per ml and incubated in 5% CO 2 at 37ЊC. Cultures were passaged every 2 to 3 days.…”

Isolation of a cell-detaching factor of Trichomonas vaginalis

“…Pentatrichomonas hominis (ATCC 3000), a human gastrointestinal trichomonad, was purchased from the American Type Culture Collection, Rockville, Md. As previously reported (11), organisms were grown in ambient air at 37°C in glass, screw-cap tubes (16 by 125 mm) containing 13 ml of Diamond TYI-S-33 medium (6) (TYI [pH 6.2]) supplemented with 10% heat-inactivated bovine serum. Cultures routinely attained a concentration of 2 x 106 to 6 x 106 T. vaginalis cells per ml if an inoculum of -3 x 103 T. vaginalis cells per tube was used.…”

Mouse intravaginal infection with Trichomonas vaginalis and role of Lactobacillus acidophilus in sustaining infection

“…Isolates of T. vaginalis were obtained from vaginal secretions from women. Organisms were grown in 5% CO2 at 37°C in glass screw-cap tubes (16 by 125 mm) held at a 450 angle in 10 ml of Diamond's TYI-S-33 medium, pH 6.2 (4), supplemented with 10% heat-inactivated fetal bovine serum and 100 U of penicillin, 100 p,g of streptomycin, and 2.5 ,ug of amphotericin B per ml as previously described (9). Cultures were passaged every 2 to 3 days.…”