A comparison of the sensitivity of the InPouch TV, Diamond's and Trichosel media for detection of Trichomonas vaginalis.

OBJECTIVE: This study compared the ability of three culture media (InPouch TV, Diamond's, and Trichosel) to support the growth of clinical isolates of Trichomonas vaginalis and their relative sensitivity for detection of the organism. METHODS: The majority of the clinical isolates were obtained from two San Francisco Bay Area clinics. T vaginalis was subcultured in 4 ml of one of the InPouch, Diamond's, or Trichosel media for 24-48 hours before evaluation. Twenty isolates were initially cultured in the InPouch test, 13 with Diamond's, and 10 with Trichosel. A haemocytometer was used to measure the initial concentrations of the organisms. Then serial dilutions were made in saline to yield approximately 2.0 x 10(4), 2.0 x 10(3), and 2.0 x 10(2) motile T vaginalis per ml. A 30 microliter inoculum from each dilution was transferred into 4 ml aliquots of the three media (387 individual tests, 43 x 3 dilutions x 3 media). Microscopic examinations for viable trichomonads were made at 24, 48, and 96 hours. Microscopy was through the pouch wall for the InPouch medium, and through a cover slipped slide with one drop of Diamond's and Trichosel media. RESULTS: At 24 hours, the InPouch demonstrated 84/129 positive, Diamond's 23/129, and Trichosel 18/129. At 48 hours, an accumulative positive rate for the InPouch was 98/129, for Diamond's 55/129, and Trichosel 47/129. At 96 hours the total positives for each test were 112/129 for the InPouch, 78/129 for Diamond's, and 74/129 for Trichosel. CONCLUSIONS: The InPouch TV test was significantly more sensitive than either Diamond's or Trichosel (at 0.01 level of significance, pInPouch > pDiamond's; pInPouch > pTrichosel on all three dilutions at 24, 48, and 96 hours). This increased sensitivity was the result of either a reduced generation time or the larger volume of media examined microscopically.

An in vitro metronidazole susceptibility test for trichomoniasis using the InPouch TV test.

OBJECTIVE: An efficient anaerobic culture system, the InPouch TV test, was used to determine the susceptibility of Trichomonas vaginalis to metronidazole. Glacial acetic acid was employed as a solvent for metronidazole. METHODS: T vaginalis isolates were cultured from 16 symptomatic female patients. The 11 who responded to oral metronidazole, 250 mg tid for 7 days, were considered as having sensitive trichomonads; the 5 who remained infected after treatment were considered to have resistant organisms. All isolates were cultured for minimum lethal concentrations (MLC). Metronidazole was added to a series of pouches; two-fold dilution of the most concentrated was 50 micrograms/ml and the least was 0.4 micrograms/ml. The inoculum of viable trichomonads was 1 x 105/ml in each pouch. Pouches were incubated at 37 degrees C for 48 h, examined microscopically for motile trichomonads, and then 0.5 ml was subcultured to drug free pouches. After 5 days incubation at 37 degrees C, each subculture and culture were examined microscopically for viable trichomonads. RESULTS: Eleven isolates of T vaginalis from patients responding to metronidazole treatment had MLC between 0.4 to 3.1 micrograms/ml. The MLC from the 5 treatment failure patients were between 12.5 to 50 micrograms/ml. CONCLUSIONS: For the 16 patients in this study, the MLC values determined with the InPouch TV test differentiated between infection caused by metronidazole sensitive and resistant trichomonads. The mean MLC of clinically resistant isolates was approximately eleven fold higher than the mean MLC of clinically sensitive isolates (15 micrograms/ml vs 1.32 micrograms/ml). There was a significant difference between clinically resistant and sensitive isolates (t = 5.47, p < 0.0005). This study suggests that the InPouch TV test could be used for distinguishing between metronidazole resistant and sensitive isolates.

Prevalence of Trichomonas vaginalis in a male sexually transmitted disease clinic population by interview, wet mount microscopy, and the InPouch TV test.

OBJECTIVE: To determine the prevalence of trichomoniasis in male patients from their urine at a Sexually Transmitted Disease (STD) Clinic using the InPouch TV culture system. METHODS: Two hundred and four patients were examined for STD infections. Their ages ranged between 17 and 72 years. Depending on their clinical symptoms tests were ordered for Neisseria gonorrhoeae, chlamydia, and for syphilis. Each patient submitted a clean catch urine specimen for trichomonas testing. A 15 ml aliquot of urine was centrifuged and a drop of the sediment examined microscopically. The remainder was cultured in the InPouch TV test. Each pouch was examined at 24 h, 48 h, and 5 days. RESULTS: Twenty-four of the 204 patients (12%) were culture positive for Trichomonas vaginalis and only three of these were wet mount positive. CONCLUSION: The InPouch TV test demonstrated an epidemiological important infected male population that was not indicated by wet mount microscopy.

Trichomoniasis: its clinical significance and diagnostic challenges.

Trichomoniasis is a significant STD worldwide. Clinical studies have demonstrated that trichomoniasis may have serious side effects both in pregnancy and in a predisposition to retrovirus infection. Therefore it is important that clinicians request the most accurate diagnostic test available. This is mandated by the significant percentage of both male and female patients that may be asymptomatic. Numerous clinical studies have indicated that the most sensitive test for T. vaginalis is with culture. The InPouch TV culture test has demonstrated high sensitivity, long shelf life, and microscopic advantages over other culture procedures.

A clinical evaluation of trichomoniasis in San Jose, Costa Rica using the InPouch TV test.

OBJECTIVE: to determine the prevalence of trichomoniasis in San Jose, Costa Rica, comparing two methods, the InPouch TV test and the saline wet mount. METHODS: One hundred symptomatic and asymptomatic female patients at two hospitals and at a sexually transmitted disease (STD) clinic were evaluated. Vaginal discharge was the most prevalent genitourinary abnormality among symptomatic patients. The patients were between 18 and 70 years old. Fifty-seven were from the STD clinic, 43 from the two hospitals. A saline wet mount and a culture were taken from each patient. The culture employed a new procedure for diagnosis of trichomonads, the InPouch TV test (BioMed Diagnostics, San Jose, CA). RESULTS: Thirteen of the 100 patients were culture positive, two of whom were wet mount positive. No wet mount positives were culture negative. Eleven of the positive tests were from the STD clinic and two were from the hospitals. CONCLUSIONS: The results of this initial epidemiologic study indicate a prevalence of 19% for trichomoniasis in the STD clinic population and 4.6% in the hospitals group. Trichomonas vaginalis was not diagnosed by laboratory methods prior to this study. The InPouch TV test has a selective fungicidal and bactericidal, enriched proteose-peptone medium which provides a sensitivity of 4 organisms per ml and a 1 year shelf life at room temperature. This in vitro culture test demonstrated unique capabilities as a transport and culture medium. Its procedure offers simplicity in application and an excellent visualisation of trichomonads.

An evaluation of an InPouch TV culture method for diagnosing Trichomonas vaginalis infection.

A new culture method for Trichomonas vaginalis, the InPouch TV test, was evaluated for its sensitivity and specificity in supporting growth of trichomonads. Five clinical isolates remained viable for periods from 41 to 131 days. A strain from the ATTC 30001 remained viable for 91 days. As few as four trichomonads/ml of culture medium could be viewed microscopically within 24 h. Doubling time for growth of trichomonad varied between 5 to 8 h. In a clinical study of 102 wet-mount negative specimens, 15 culture positive patients were observed with the InPouch TV test compared with 12 of the same patients with Hollander's fluid medium.

The effects of 3-deazaguanine on Rous sarcoma virus transformation.

An expression of cellular transformation in chicken embryo fibroblast infected with Rous sarcoma virus (RSV) was inhibited by 3-deazaguanine (3-DG). Re-transformation of normal rat kidney cells infected with a temperature-sensitive mutant of RSV was suppressed by 3-DG when the cells were shifted from the non-permissive temperature (38 degrees C) to the permissive temperature (33 degrees C). Removal of the inhibitor enabled the cells to revert to the transformed state. Finally, 3-DG had little or no effect on replication of RSV in the chicken embryo fibroblast cells.

Morphological aspects of inhibition of transformation of rat kidney cells infected with a temperature-sensitive mutant of the Rous sarcoma virus by rat fibroblast interferon.

Normal rat kidney cells (NRK) infected with a temperature-sensitive (ts) mutant of Rous Sarcoma virus (RSV) at the permissive temperature (33 degrees C) exhibit the morphological and growth characteristics of the transformed state. By the use of this system, it has been shown that the addition of rat fibroblast interferon to infected cells grown at a nonpermissive temperature (38 degrees C) prevents re-transformation when they are introduced to the permissive temperature. Merely treating cells at the permissive temperature with rat fibroblast interferon causes them to change from the transformed phenotype to a normal phenotype. These effects require the continuous presence of interferon and are quite species specific as observed with the ineffectiveness of mouse fibroblast interferon. Therefore, the results suggest that interferon might restore the growth control of transformed cells which could in part accounts for its anti-tumor effect.

A 24 hour plastic envelope method for isolating and identifying Gardnerella vaginalis (PEM-GVA)

A new plastic envelope culture test that is selective for Gardnerella vaginalis was compared with a conventional method. Vaginal specimens from 92 women were cultured. Results from both methods were compared with the results of pelvic examinations and clinic screening tests used to diagnose bacterial vaginosis (BV). G vaginalis was isolated more often in the envelope than by the conventional method from patients with BV and those without, though the difference was not significant. Isolation and identification of G vaginalis was completed in 18-24 hours by the envelope method; the conventional method took a mean of 72 hours (range two to six days). Polymorphonuclear leucocytes (PMNLs) occurred significantly more in specimens from the patients without BV than from those with BV. Both clue cells and a positive amine test reaction were found significantly more in specimens from patients with BV than from those without BV. Clue cells and G vaginalis isolation correlated best with BV (in 47 women), followed by clue cells and positive amine test results (in 39). Adherence of G vaginalis in the envelope also correlated more with BV, clue cells, and positive amine test results (32) than with patients without BV (14). When there were no clue cells and amine test results were negative the results correlated totally with a prediction of no BV. The use of the rapid envelope culture test would have confirmed BV in 20% of the cases where clue cell and amine test results were discordant.

Enteric pathogenic protozoa in homosexual men from San Francisco.

The prevalence of enteric protozoa was studied in a survey conducted among 150 male homosexual patients in San Francisco. All patients were from a private practice in internal medicine. Each was asked to complete a questionnaire and to submit multiple stool specimens for examination. Of this group, 47% were positive for one or more potentially pathogenic intestinal protozoa; Entamoeba histolytica was found in 36%, Entamoeba hartmanni in 35%, Giardia lamblia in 5%, and Dientamoeba fragilis in 1.3%. Symptoms were unreliable as a diagnostic index of intestinal protozoan infection. Colonization rates could not be correlated with any specific sexual technique. The large number of homosexuals at risk, combined with the potential for difficulties in diagnosis and contact-tracing, indicate the possibility that enteric pathogenic protozoa will cause future health problems in this population.

Localization of catheter-induced urinary tract infections: interpretation of bladder washout and antibody-coated bacteria tests.

Accurate differentiation of upper from lower urinary tract infection (UTI) has been reported based on the presence or absence of antibody-coated bacteria (ACB) in the urinary sediment. The specificity of ACB localization in adults has been reported to be high based on a strong correlation with localization results obtained with the bladder washout technique. The present study examines the efficacy of ACB testing with respect to accurate anatomic localization of UTI in 20 studies in 14 patients exhibiting bacteriuria as a result of chronic urethral catheterization. 17 studies (85%) demonstrated significant ACB. Localization of UTI was assessed in each study by the bladder washout technique. Following instillation of a washout solution, a normal decrease in colony counts was observed in only 3 studies (15%), a frequency significantly lower than the value of 90% previously reported in noncatheterized bacteriuric patients (p less than 0.001). Each of these 3 patients was ACB-positive, yet anatomic localization indicated lower tract UTI. The observed characteristics of the 14 patients included the absence of upper tract signs and symptoms, and spontaneous clearing of ACB and bacteriuria following catheter removal. These findings suggest that in chronically catheterized patients, both (1) the observed abnormal resistance of bacteria to bladder washout and (2) the high frequency of positive ACB tests are indices of severe chronic tissue invasion and intense antibody production in the lower urinary tract in this condition and thus not of value in localizing infection to the upper tract. Neither ACB nor washout testing with current techniques can reliably localize UTI in chronically catheterized patients.

Intestinal parasites in Southeast Asian refugees.

A survey of intestinal parasitism in 6,241 Southeast Asian refugees (3,576 males and 2,665 females) indicated that 32.9 percent of the refugees had one or more intestinal parasites. A total of 1,178 (57.3 percent) males and 878 (42.7 percent) females harbored the parasites, with helminths representing the most frequent isolate. Intestinal parasitic infections may be considered minimal public health threats in the United States because of effective hygienic practices and sanitation facilities. However, it is important to emphasize that the attack rate also will be influenced by continued health education, job activities, and diagnosis and treatment of the refugees for these parasites.

Micromethod for rapid identification of gram-negative, nonfermentative bacteria.

Because of the contemporary significance of gram-negative, nonfermentative bacteria as etiological agents, a simple, rapid determinative system is essential. Therefore, an accurate, reproducible, and an inexpensive micromethod for the identification of these organisms has been developed. Included in this system are twenty-five biochemical tests. Carbohydrate utilization is demonstrated by modification of Otto and Pickett's formula for oxidative attack and assimilation of carbon sources, while the other substrates are modifications of commercially available products. Inoculation is a two-fold procedure into a plastic multicompartmented tray with wells containing 100 micro liters of each substrate. Initial inoculation yields 10(5) colony forming units per well. The carbohydrates are supplemented with an additional 50 micro liters of a 1 X 10(11) saline suspension of organisms. Reactions are read after a maximum incubation of 48 hr at 35 C. The results obtained with the identification of 124 strains of nonfermentative bacteria utilizing a conventional media schema as suggested by the Center for Disease Control and the micromethod system demonstrated the high reproducibility and correlation achievable with this rapid and economical microtechnique.