Comparison of Resistant and Susceptible Strains of Trichomons vaginalis to Metronidazole Using PCR Method
,
Abstract
Background: 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.
Methods: 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.
Results: 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 µg/ml; however, this concentration for resistant isolates was 200 µg/ml after 24 h and 100 µg/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.
Conclusion: Resistance to metronidazole in T. vaginalis has not relation to genetic variations and might be related to some physiologic pathways of organism.
Meri T, Jokiranta TS, Suhonen L, Meri S. Resistance of Trichomonas vaginalis to metronidazole: report of the first three cases from Finland and optimization of in vitro susceptibility testing under various oxygen concentrations. J Clin Microbiol. 2000; 38(2):763-7.
Snipes LJ, Gamard PM, Narcisi EM, Beard CB, Lehmann T, Secor WE. Molecular epidemiology of metronidazole resistance in a population of Trichomonas vaginalis clinical isolates. J Clin Microbiol. 2000; 38(8):3004-3009.
Johanson S, Sanchez JL, Gerding DN. Metronidazole resistance in Clostridium difficile. Clin Infect Dis. 2000; 31:625-6.
Löfmark S, Edlund C, Nord CE. Metronidazole is still the drug of choice for treatment of anaerobic infections. Clin Infect Dis. 2010; 1;50 Suppl 1:S16-23.
Schmidt G, Narcici E, Mosure D, Secor WE, Higgins J, Moreno H. Prevalence of metronidazole resistant Trichomonas vaginalis in a gynecology clinic. J Reprod Med. 2001; 46: 545-549.
Schwebke JR, Barrientes FJ, 2006. Prevalence of Trichomonas vaginalis isolates with resistance to metronidazole and tinidazole. Antimicrob Agents Chemother. 50(12): 4209-10.
Perez S, Fernandez-Verdugo A, Perez F, Vasquez F. Prevalence of 5-nitroimidazole resistant Trichomonas vaginalis in Oviedo, Spain. Sex Transm Dis. 2001; 28: 115-16
Schwebke JR, Burgess D. Tric-homoniasis. Clin Microbiol Rev. 2004; 17:794-803.
Rosolson M, Vanacova S, Tomkova E, Rezga J, Hardy IJ, Kulda J. Mechanisms of in vitro development of resistance to metronidazole in Trichomonas vaginalis. Microbiology. 2002; 148: 2467-2477.
Narcisi EM, Secor WE. In vitro effect of tinidazole and furazolidone on metronidazole- resistant Trichomonas vaginalis. Antimicrob. Agents Chemother. 1996; 40:1121-1125.
Upcroft P, Upcroft JA. Drug susceptibility testing of anaerobic protozoa. Antimicrobial Agents and Chemotherapy. 2001; 45(6):1810-1814.
Pillay A, Lewis J, Ballard RC. Evaluation of Xenostrip-Tv, a rapid diagnostic test for Trichomonas vaginalis infection. J Clin Microbiol. 2004; 42(8): 3853-3856.
Ali V, Nozaki T. Current therapeutics, their problems, and sulfur-containing-amino-acid metabolism as a novel target against infections by "Amithochondriate" protozoan parasites. Clin Microbiol Rev. 2007; 20(1): 164-87.
Land KM, Clemens DL, Johnson PJ. Loss of multiple hydrogenosomal proteins associated with organelle metabolism and high-level drug resistance in trichomonads. Exp Parasitol. 2001; 97:102-110.
Rasoloson DE, Tomkova R, Cammack J et al. Metronidazole-resistant strains of Trichomonas vaginalis display increased susceptibility to oxygen. Parasitology. 2001; 123:45-56.
Land KM, Johnson PJ. Molecular basis of metronidazole resistance in pathogenic bacteria and protozoa. Drug Resist Updat. 1999; 2:289-94.
Theron MM, Jonse Van Rensburg MN, Chelkley Lj. Nitroimidazole resistance genes (nimB) in anaerobic Gram-positive cocci (previously Peptostreptococcus spp.). J Antimicrob Chemother. 2004; 54:240-2.
Lofmark S, Fang H, Hedberg M, Edlund C. Inducible metronidazole resistance and nim genes in clinical Bacteroides fragilis group isolates. Antimicrob Agents Chemother. 2005; 49: 1253-6.
Whittle G, Shoemaker NB, Salyers AA. The role of Bacteroides conjugative transposons in the dissemination of antibiotic resistance genes. Cell Mol Life Sci. 2002; 59: 2044-54.
Vatanshenassan M, Rezaie S, Mohebali M, Niromand N, Kazemi B, Babaei Z, Rezaeian M. Trichomonas vaginalis: investigation of a novel diagnostic method in urine samples using cysteine proteinase 4 gene and PCR technique. Exp Parasitol. 2010; 126(2):187-90.
Rabiee S, Fallah M, Zahabi F. Frequency of Trichomoniasis in patients admitted to outpatient clinics in Hamadan (2007) and relationship between clinical diagnosis and laboratory findings. J Res Health Sci. 2010; 9(1): 31-35.
Rezaeian M, Vatanshenassan m, Rezaie S, Mohebali M, Niromand N, Niyyati M, Farnia S, Babaei Z. Prevalence of Trichomonas vaginalis Using Parasitological Methods in Tehran. Iranian J Parasitol. 2009; 4(4): 43-47.
Maraghi S, Khosravi A, Kardooni T, T Razi T, Feiz-Haddad MH. Evaluation of an Immunochromatographic Strip (Xenostrip - Tv) test for diagnosis of vaginal trichomoniasis compared with wet mount and PCR assay. Iranian J Parasitol. 2008; 3(3) : 11-17
Valadkhani Z, Assmar M, N. Hassan N, Aghighi Z, Amirkhani A, Kazemi F, Esmaili I, Samandar M, Moradynasab M, Sabzali D. The prevalence of trichomoniasis in high-risk behavior group women attending penitentiaries clinic of Tehran province. Giornale Italiano di Medicina Tropicale. 2009; 14(1-4): 43-46
25 Kazemi F, Hooshyar H, Zareikar B, Bandehpour B, Arbabi M, Talari S, Alizadeh R, Kazemi B. Study on ITS1 Gene of Iranian Trichomonas vaginalis by Molecular Methods. Iranian J Parasitol. 2010; 5(4): 9-14.
Valadkhani Z, Kazemi F, Hassan N, Aghighi Z, Esmaili I, Talebi M. Gene Diversity of Trichomonas vaginalis Isolates. Iranian J Parasitol. 2011; 6 (3):101-106.
Jamali R, Zareikar B, Kazemi A et al. Random amplified polymorphic DNA of Trichomonas vaginalis isolates from Tabriz, Iran. J Parasitic Dis. 2006; 30 (1): 53-57.
Blaha C, Duchêne M, Aspöck H, Walochnik J. In vitro activity of hex-adecylphosphocholine (miltefosine) against metronidazoleresistant and susceptible strains of Trichomonas vaginalis. J Antimicrob Chemother. 2006; 57(2):273-8.
| Files | ||
| Issue | Vol 7 No 3 (2012) | |
| Section | Articles | |
| Keywords | ||
| Trichomonas vaginalis Metronidazole Resistance PCR | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
