Nucleic Acid Testing for Trichomonas vaginalis

2016, Volume 26, Number 1

Author

Danny Wiedbrauk, Ph.D.,
Scientific Director, Virology and Molecular Biology, Warde Medical Laboratory


Trichomonas vaginalis affects men and womenTrichomoniasis is a persistent disease of the genitourinary tract caused by the flagellated protozoan Trichomonas vaginalis. Trichomoniasis is the most common nonviral sexually transmitted disease (STD) in the United States, affecting an estimated 3.7 million people (1). The overall prevalence of T. vaginalis infection is >11% in women aged ≥40 years (2). In symptomatic women who visit STD clinics, the reported prevalence is 26% (3) and in one study of incarcerated individuals, the infection rates were 9%–32% for women and 2-9% for men.

Symptoms

Seventy to eighty-five percent of T. vaginalis– infected individuals have minimal or no symptoms and when left untreated, these infections can last for months to years. (4-7) Symptomatic women can present with vaginitis with small petechial or sometimes punctate red “strawberry” spots and profuse, thin, foamy, greenish-yellow discharge with foul odor. The disease may also cause cystitis or urethritis.  Vulvar involvement is variable. In men, T. vaginalis can cause urethritis, epididymitis, or prostatitis. These infections cause 5-10% of non-gonococcal urethritis in men. (8)

Complications

Trichomonas vaginalis infection is associated with adverse pregnancy outcomes including premature rupture of membranes, preterm delivery, and delivery of a low birthweight infant. (9-12) Infection is also associated with two- to threefold increased risk for HIV acquisition. (13-16) Among women with HIV, T. vaginalis infection is associated with increased risk for pelvic inflammatory disease (17-19) and routine screening of asymptomatic women with HIV infection is recommended because of the adverse events associated with trichomoniasis and HIV infection.

Transmission/Prevention

Although partners might be unaware of their infection, organisms are readily passed between sex partners during penile-vaginal sex. (8)  Partners of men who have been circumcised might have a somewhat reduced risk of T. vaginalis infection acquisition. (20, 21) There is no vaccine for trichomoniasis, and for sexually active individuals, the best way to prevent trichomoniasis is through consistent and correct use of condoms during all penile-vaginal sexual encounters. (22)

Persistent or Recurrent Infection

While most recurrent T. vaginalis infections are thought to result from reinfection, some infections can be attributed to antimicrobial resistance. Metronidazole resistance occurs in 4%–10% of cases of vaginal trichomoniasis (23, 24)  and tinidazole resistance in 1%. (24) Emerging nitroimidazole-resistant trichomoniasis is concerning because few alternatives to standard therapy exist. The Centers for Disease Control and Infection (CDC) has experience with susceptibility testing for nitroimidazole-resistant T. vaginalis and can provide Nucleic Acid Testing for Trichomonas vaginalis treatment assistance in these cases (telephone: 404-718-4141; website: http://www.cdc.gov/std)

holding hands: T. vaginalis infections are STDsWho Should be Tested?

As mentioned previously, the CDC recommends routine screening of asymptomatic women with HIV infection because of the adverse events associated with trichomoniasis and HIV infection.  Diagnostic testing should also be performed in women seeking care for vaginal discharge.

Screening might be considered for persons receiving care in high-prevalence settings (e.g., STD clinics and correctional facilities) and for asymptomatic persons at high risk for infection (e.g., persons with multiple sex partners, exchanging sex for payment, illicit drug use, or a history of STD).

However, data are lacking on whether screening and treatment for asymptomatic trichomoniasis in high prevalence settings or persons at high risk can reduce any adverse health events and health disparities or reduce community burden of infection. (8)  Rectal and oral testing for T.vaginalis is not recommended. (8)

Testing Methods

MICROSCOPIC EVALUATION

Microscopic evaluation of wet mount preparations is the most common method for the diagnosis of T. vaginalis genital infections due to the convenience of the procedure and its relatively low cost. Unfortunately, the sensitivity of the wet mount procedure is low (51-65%) for vaginal specimens (Table 1) and lower still in specimens from men (urethral specimens, urine sediments, and semen. (25) Evaluation of wet mount preps must be done promptly after collection because the already low sensitivity of the procedure declines by to 20% within one hour after collection. (26, 27) While T. vaginalis may be an incidental finding in a Pap test, microscopic examination of conventional or liquid-based Pap specimens should not be used as the primary means for detecting T. vaginalis because false positive and false negative results can occur. (8)

CULTURE

Culture was considered the gold standard method for diagnosing T. vaginalis infection before molecular detection methods became available. Culture has a sensitivity of 75%–96% and a specificity of up to 100%. (25)  In women, vaginal secretions are the preferred specimen type for culture, as urine culture is less sensitive. (25, 28, 29) In men, culture specimens require a urethral swab, urine sediment, and/or semen. To improve yield, multiple specimens from men can be used to inoculate a single culture.

NUCLEIC ACID AMPLIFICATION TESTING (NAAT)

Nucleic acid amplification testing is rapidly replacing culture as the gold-standard for T. vaginalis testing. In the 2015 STD treatment guidelines (8), the CDC recommends the use of highly sensitive NAAT testing for the detection of T. vaginalis. Among women, NAAT detects three to five times more T. vaginalis infections than wet-mount microscopy. (30, 31) When NAAT testing on specimens is not feasible, a testing algorithm (e.g., wet mount first, followed by NAAT if negative) can improve diagnostic sensitivity in persons with an initial negative result by wet mount. (25)

Table 1. Sensitivity of diagnostic methods for detecting
Trichomonas vaginalis in vaginal specimens.
Test MethodSensitivityTime to Result
Wet Mount51-65%<1 hour
Culture75-96%1-4 days
Nucleic Acids95-100%2-3 days

Trichomonas vaginalis testing at Warde

Warde Medical Laboratory, uses the highly sensitive and specific APTIMA Trichomonas vaginalis Assay for the detection of T. vaginalis ribosomal RNA in clinician-collected endocervical swabs, clinician- collected vaginal swabs, female first-void urine specimens, and specimens in PreservCyt Solution.

Specimens can be collected from symptomatic and asymptomatic patients.  We have also validated the use of male first-void urine specimens and male urethral specimens with this test.  The overall sensitivity of the assay is 95.2% and the specificity is 98.0%.  The analytical sensitivity of the APTIMA procedure is 0.1 organism/mL.

Clinical testing is performed Monday – Friday and the time to result is 1-3 days.  More information about specimen stability, test codes, and LOINC Codes can be found in the Test Catalog on the Warde Medical Laboratory website (wardelab.com).

Literature Cited
  1. Sutton M, Sternberg M, Koumans EH, et al. The prevalence of Trichomonas vaginalis infection among reproductive-age women in the United States, 2001-2004. Clin Infect Dis 2007;45:1319–26.
  2. Ginocchio CC, Chapin K, Smith JS, et al. Prevalence of Trichomonas vaginalis and coinfection with Chlamydia trachomatis and Neisseria gonorrhoeae in the United States as determined by the Aptima Trichomonas vaginalis nucleic acid amplification assay. J Clin Microbiol 2012;50:2601–8.
  3. Meites E, Llata E, Braxton J, et al. Trichomonas vaginalis in selected U.S. sexually transmitted disease clinics: testing, screening, and prevalence. Sex Transm Dis 2013;40:865–9.
  4. Peterman TA, Tian LH, Metcalf CA, et al. High incidence of new sexually transmitted infections in the year following a sexually transmitted infection: a case for rescreening. Ann Intern Med 2006;145:564–72.
  5. Sutton M, Sternberg M, Koumans EH, et al. The prevalence of Trichomonas vaginalis infection among reproductive-age women in the United States, 2001-2004. Clin Infect Dis 2007;45:1319–26.
  6. Peterman TA, Tian LH, Metcalf CA, et al. Persistent, undetected Trichomonas vaginalis infections? Clin Infect Dis 2009;48:259–60.
  7. Gatski M, Kissinger P. Observation of probable persistent, undetected Trichomonas vaginalis infection among HIV-positive women. Clin Infect Dis 2010;51:114–5.
  8. CDC. Centers for Disease Control and Prevention. Sexually Transmitted Diseases Treatment Guidelines, 2015. MMWR Recomm Rep 2015;64(No. RR-3): 1-137.
  9. Cotch MF, Pastorek JG, 2nd, Nugent RP, et al. Trichomonas vaginalis associated with low birth weight and preterm delivery. Sex Transm Dis 1997;24:353–60.
  10. Mann JR, McDermott S, Gill T. Sexually transmitted infection is associated with increased risk of preterm birth in South Carolina women insured by Medicaid. J Matern Fetal Neonatal Med 2010;23:563–8.
  11. Mann JR, McDermott S, Barnes TL, et al. Trichomoniasis in pregnancy and mental retardation in children. Ann Epidemiol 2009;19:891–9.
  12. Mann JR, McDermott S. Are maternal genitourinary infection and pre-eclampsia associated with ADHD in school-aged children?J Atten Disord 2011;15:667–73.
  13. McClelland RS, Sangare L, Hassan WM, et al. Infection with Trichomonas vaginalis increases the risk of HIV-1 acquisition. J Infect Dis 2007;195:698–702.
  14. Van Der Pol B, Kwok C, Pierre-Louis B, et al. Trichomonas vaginalis infection and human immunodeficiency virus acquisition in African women. J Infect Dis 2008;197:548–54.
  15. Hughes JP, Baeten JM, Lingappa JR, et al. Determinants of per-coital-act HIV-1 infectivity among African HIV-1-serodiscordant couples. J Infect Dis 2012;205:358–65.
  16. Kissinger P, Adamski A. Trichomoniasis and HIV interactions: a review. Sex Transm Infect 2013;89:426–33.
  17. 17. Minkoff H, Grunebaum AN, Schwarz RH, et al. Risk factors for prematurity and premature rupture of membranes: a prospective study of the vaginal flora in pregnancy. Am J Obstet Gynecol 1984;150:965–72.
  18. Cotch MF, Pastorek JG, 2nd, Nugent RP, et al. Trichomonas vaginalis associated with low birth weight and preterm delivery. Sex Transm Dis 1997;24:353–60.
  19. Moodley P, Wilkinson D, Connolly C, et al. Trichomonas vaginalis is associated with pelvic inflammatory disease in women infected with human immunodeficiency virus.Clin Infect Dis 2002;34:519–22.
  20. Sobngwi-Tambekou J, Taljaard D, Nieuwoudt M, et al. Male circumcision and Neisseria gonorrhoeae, Chlamydia trachomatis and Trichomonas vaginalis: observations after a randomised controlled trial for HIV prevention. Sex Transm Infect 2009;85:116–20.
  21. Gray RH, Kigozi G, Serwadda D, et al. The effects of male circumcision on female partners' genital tract symptoms and vaginal infections in a randomized trial in Rakai, Uganda. Am J Obstet Gynecol 2009;200:e41–7.
  22. Crosby RA, Charnigo RA, Weathers C, et al. Condom effectiveness against non-viral sexually transmitted infections: a prospective study using electronic daily diaries. Sex Transm Infect 2012;88:484–9.
  23. Kirkcaldy RD, Augostini P, Asbel LE, et al. Trichomonas vaginalis antimicrobial drug resistance in 6 US cities, STD Surveillance Network, 2009-2010. Emerg Infect Dis 2012;18:939–43.
  24. Schwebke JR, Barrientes FJ. Prevalence of Trichomonas vaginalis isolates with resistance to metronidazole and tinidazole. Antimicrob Agents Chemother 2006;50:4209–10.
  25. Nye MB, Schwebke JR, Body BA. Comparison of APTIMA Trichomonas vaginalis transcription- mediated amplification to wet mount microscopy, culture, and polymerase chain reaction for diagnosis of trichomoniasis in men and women.Am J Obstet Gynecol 2009;200:e181–7.
  26. Stoner KA, Rabe LK, Meyn LA, et al. Survival of Trichomonas vaginalis in wet preparation and on wet mount.Sex Transm Infect 2013;89:485–8.
  27. Kingston MA, Bansal D, Carlin EM. 'Shelf life' of Trichomonas vaginalis. International journal of STD and AIDS2003;14:28–9.
  28. Mohamed OA, Cohen CR, Kungu D, et al. Urine proves a poor specimen for culture of Trichomonas vaginalis in women. Sex Transm Infect 2001;77:78–9.
  29. Lawing LF, Hedges SR, Schwebke JR. Detection of trichomonosis in vaginal and urine specimens from women by culture and PCR. J Clin Microbiol 2000;38:3585–8.
  30. Hollman D, Coupey SM, Fox AS, et al. Screening for Trichomonas vaginalis in high-risk adolescent females with a new transcription-mediated nucleic acid amplification test (NAAT): associations with ethnicity, symptoms, and prior and current STIs. J Pediatr Adolesc Gynecol 2010;23:312–6.
  31. Roth AM, Williams JA, Ly R, et al. Changing sexually transmitted infection screening protocol will result in improved case finding for Trichomonas vaginalis among high-risk female populations. Sex Transm Dis 2011;38:398–400.