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The impact of the viability of Trichomonas vaginalis in urine on wet mount, culture, and PCR methods was assessed. To minimize the chance of false-negative results, urine specimens should be processed within 30 min of specimen collection and maintained at 37 degrees C, since temperature appears to affect the viability of Trichomonas.
Antibodies against Trichomonas vaginalis were detected in serum samples from 98 patients by three immunological assays. A good correlation was observed between the enzyme-linked immunosorbent assay and the immunofluorescence method, whereas it was found that the enzyme-linked immunosorbent assay correlated better with the current or past detection of organisms than did other serological methods (immunofluorescence and hemagglutination). Similar results were obtained with whole trichomonads and two T. vaginalis soluble antigenic preparations, which suggests that immunodominant moieties shared by several T. vaginalis strains were detected. The level of antibodies of the immunoglobulin A class was higher in patients with past records of trichomoniasis, but less significantly so than the total antitrichomonal antibody level. Antibodies of the four immunoglobulin G subclasses were detected. Immunoglobulin G1 antibody values were higher in female than male patients.
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 10(6) to 6 X 10(6) 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 10(6) to 6 X 10(6) T. vaginalis per ml, but at least 3 X 10(3) 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. 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 women. 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 numbers.
We developed a polymerase chain reaction (PCR)-based test for detecting the protozoan parasite Trichomonas vaginalis. Genomic libraries were constructed from two independent clinical isolates of T. vaginalis. From these libraries, 12 genomic clones were purified, sequenced, and then screened for uniqueness by computer-assisted sequence comparisons. PCR reactions were performed to evaluate eight PCR-primer pairs, including a primer pair that targeted the T. vaginalis ferredoxin gene. All eight primer pairs yielded PCR products of the expected sizes. However, six of the primer pairs amplified their respective target sequences in limited numbers of clinical T. vaginalis isolates, suggesting the presence of significant genomic variability among isolates. An exception was a primer pair, termed TVA5-TVA6, that amplified a 102-bp genomic sequence, termed A6p, in all of 24 clinical isolates. The A6p sequence was not detected by PCR in human DNA or in a wide variety of flagellates, ciliates, or bacteria tested. The A6p sequence appears highly selective for a broad range of T. vaginalis isolates and holds promise for PCR-based diagnosis of the parasite.
Although culture of Trichomonas vaginalis is more sensitive than wet mounts in the diagnosis of trichomoniasis, the lack of convenience of culture prevents it from being widely used. To improve the acceptability of diagnosis by culture, a plastic envelope method (PEM) was devised. PEM permits both immediate examination and culture in one self-contained system. The medium consists of dry ingredients that are reconstituted with water before use. The effectiveness of immediate examinations by PEM was compared with that of wet mounts, and the effectiveness of culture by PEM was compared with that of culture in Trichomonas Medium No. 2 (Oxoid). Of 710 vaginal secretion specimens from symptomatic and asymptomatic women that were tested by the four methods, 62 (9%) were positive for T. vaginalis. The sensitivity was 66% by wet mount, 66% by immediate examination by PEM, 89% by cultures in Oxoid medium, and 97% by culture by PEM. The two culture methods had equivalent sensitivities but were significantly (P less than 0.0001) more sensitive than the two immediate methods. The combined immediate examination by PEM plus culture was more convenient to use than wet mounts plus culture in Oxoid medium. The long shelf-life of PEM's dry medium and its anticipated low cost are additional advantages.
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