Detection of Trichomonaisis in Vaginal and Urine Specimens from Women by Culture and PCR

1. Detection of Trichomonaisis in Vaginal and Urine Specimensfrom Women by Culture and PCR Mariska Petersen

2. Trichomonaisis • History • Biological Mechanism • Experimentation • Results

3. Trichomonaisis • Sexually transmitted infection • Discovered in 1836 • 5 million cases reported every year • Resides in genitourinary tract of males and females • 50% asymptomatic carriers

4. Transmission • Sexual intercourse • Mutual masturbation • Sharing sex toys • Mother to child during vaginal delivery

5. In females there is… inflammation of the vaginal canal vulvar itching leading to edema tenderness and chaffing redness yellow and green discharge painful urination pain during sex punctuate hemorrhages on the cervix known as colpitis macularis or strawberry cervix. In males there is… whitish discharge although rare pain and burning during urination Signs and Symptoms

6. Treatment Metronidazole (Flagyl) ~single two gram dose (85-92% cure rate) OR 250 mg three time daily for 7-10 days. Tinidazole is an alternate drug

7. Biological Mechanism • T. vaginalis – causitive agent • Reproduction- binary fission • Incubation period-4-28 days • Facultative anerobic parasite • Adhesion proteins on surface of flagella • Ligand/Receptor Cytoadherence

8. Experiment • Screening for Trichomonas vaginalis by wet-preparation microscopy and culture. • Presence of T. vaginalis DNA by specific PCR of vaginal and urine specimens.

9. Wet preparation microscopy- • PCR-

10. Materials and Methods • Female patients from the Jefferson County Department of Health STD clinic were screened for trichomonaisis. • Routine pelvic examinations were performed • Swab specimens collected • Urine samples examined

11. Initial Testing • 1st swab was used to inoculate culture medium for T. vaginalis • 2nd swab placed in cryogenic airtight vial for PCR studies • Wet preps were examined as a part of the routine exam • All vaginal symptoms were recorded • Urine samples were pelleted () at 1000 g for 5 minutes, decanted and re-suspended in 250 mL of sterile water. • Culture were examined daily for 5 days for the presence of motile trichomonads.

12. PCR Testing • Vaginal swabs agitated in 1mL of water then centrifuged at 2,000 3 g for 10 min. • DNA extracted

13. Results • Samples were recorded from 190 women • Overall presence of trichomonas was 28% • Culture of vaginal fluid most sensitive method for the detection of T. vaginalis- 94.3% • Direct microscopy least sensitive-58.5% • PCR-based detection of T. vaginalis from vaginal swabs was equivalent to culture

15. Results cont’d • In 74.5% of women motile trichomonads were detected in urine samples. • PCR for the urine specimens detected 68.4% of motile trichomonads. • 75% of women with trichomonads complained of symptoms including discharge, odor and itching.

17. Conclusion • T. vaginalis was detected in the urine of 75% of women with trichomoniasis using standard methods • PCR of urine for T. vaginalis had a sensitivity less than that of microscopy and culture • PCR for T. vaginalis using vaginal swab specimens was equivalent to culture

18. References

19. Questions???