Comparison of resistant and susceptible strains of Trichomons vaginalis to metronidazole using PCR method

ABSTRACT

Metronidazole is drug of choice recommended by WHO for treatment of trichomoniasis, however, some reports claims drug resistance in Trichomonas vaginalis isolates recently. The objective of this study was to determine the minimum lethal concentration [MLC] of metronidazole in resistant and sensitive strains, as well as genetic patterns of these stains by PCR method. From February 2006 to March 2007, in a cross sectional study, clinical and wet mount examination of vaginal smear along with culture were performed on 683 women attending to public and private outpatient clinics in Hamadan. Trichomoniasis marked based on major clinical symptoms. Diagnosis confirmed using wet mount microscopically and culture in Diamond medium. A serial concentration of metronidazole was provided and all isolated Trichomonas strains [resistant and sensitive] tested by standard method. Finally, all sensitive and resistant strains examined by PCR technique. Only 15/683, [2.2%] of patients clinically diagnosed trichomonal vaginitis were positive for T. vaginalis by wet smear and culture. The minimum lethal concentration [MLC] for clinically sensitive isolates was 25 micro g/ml; however, this concentration for resistant isolates was 200 micro g/ml after 24 h and 100 micro g/ml after 50 h. The results of PCR examination of DNA from sensitive and resistant isolates had same pattern. The lanes appeared by two primers were 98 bp and 261 bp for both clinically sensitive and resistant strains. Resistance to metronidazole in T. vaginalis has not relation to genetic variations and might be related to some physiologic pathways of organism