Molecular Characterization of Trichomonas vaginalis Strains Based on Identifying Their Probable Variations in Asymptomatic Patients

  • Adel SPOTIN Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Department of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
  • Sanaz TAGHIZADEH EGHTEDAR Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Department of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
  • Abbas SHAHBAZI Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Department of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
  • Asghar SALEHPOUR Department of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
  • Seddigheh SARAFRAZ Department of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran East Azerbaijan Province Health Center, Tabriz Health Center, Tabriz, Iran
  • Seyyed Ali SHARIATZADEH Department of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
  • Mahmoud MAHAMI-OSKOUEI Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Department of Parasitology and Mycology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
Keywords: Trichomonas vaginalis, Actin gene, Genotypes, Asymptomatic infection

Abstract

Background: The aim of this study was to identify the Trichomonas vaginalis strains/haplotypes based on identifying their probable variations in asymptomatic patients referred to Tabriz health centers, northwestern Iran.Methods: Sampling was taken from 50-suspected women to T. vaginalis in northwestern Iran. The obtained samples were smeared and cultured. Fifty DNA samples were extracted, amplified and identified by nested polymerase chain reaction and PCR-RFLP of actin gene using two endonuclease enzymes: MseI and RsaI. To reconfirm, the amplicons of actin gene were directly sequenced in order to identify the strains/haplotypes.Results: PCR-RFLP patterns, sequencing and phylogenetic analyses revealed definitely the presence of the G (n=22; 73.4%) and E (n=8; 26.6%) strains. Multiple alignments findings of genotype G showed five haplotypes and two amino acid substitutions in codons 192 and 211 although, no remarkable unique haplotype was found in genotype E.Conclusion: The accurate identification of T. vaginalis strains based on discrimination of their unknown haplotypes particularly those which are impacted on protein translation should be considered in parasite status, drug resistance, mixed infection with HIV and monitoring of asymptomatic trichomoniasis in the region.

References

World Health Organization. Global prevalence and incidence of selected curable sexually transmitted infections: overview and estimates. Geneva, Switzerland: WHO, 2001.

Fouts A, Kraus SJ. Trichomonas vaginalis: re evaluation of its clinical presentation and laboratory diagnosis. J Infect Dis. 1980; 141: 137–143.

Cates W, Joesoef RJ, Goldman M. Atypical pelvic inflammatory disease: can we identify clinical predictors? Am J Obstet Gynecol. 1993; 169: 341–346.

Cotch MF, Pastorek JG, Nugent RP et al. Trichomonas vaginalis associated with low birth weight and pre-term delivery. Sex Transm Infect 1997; 24: 353–360.

Sutton MY, Sternberg M, Nsuami M, Behets F, Nelson AM, St Louis M. Trichomoniasis in pregnant human immunodeficiency virus-infected and human immunodeficiency virus-uninfected Congolese women: prevalence, risk factors, and association with low birth weight. Am J Obstet Gynecol. 1999; 181: 656–662.

Petrin D, Delgaty K, Gaiber Bhatt R, Garber G. Clinical and microbiological aspects of Trichomonas vaginalis. Clin Microbiol Rev. 1998; 11:300 –17.

World Health Organization. An overview of selected curable sexually transmitted diseases. In: World Health Organization editor. Global programme on AIDS. Geneva, Switzerland: World Health Organization, 1995.

Alderete JF, Kasmala L, Metcalfe E, Garza GE. Phenotypic variation and diversity among Trichomonas vaginalis isolates and correlation of phenotype with trichomonal virulence determinants. Infect lmmun. 1986; 53:285 –93.

Nash TE, McCutchan T, Keister D, Dame JB, Conrad JD, Gilin FD. Restriction endonuclease analysis of DNA from 15 Giardia isolates obtained from humans and animals. J Infect Dis. 1985; 152:64 –73.

Singh B. Molecular methods for diagnosis and epidemiological studies of parasitic infections. Int J Parasitol. 1997; 27: 1135–1145.

Vanacova S, Tachezy J, Kulda J, Flegr J. Characterization of trichomonad species and strains by PCR fingerprinting. J Euk Microbiol. 1997; 44: 545–552.

Stiles JK, Shar PH, Xue L, Meade JC, Lushbaugh WB, Cleary JD, Finley RW. Molecular typing of Trichomonas vaginalis isolates by HSP70 restriction fragment length polymorphism. Am J Med Hyg. 2000; 62: 441–445.

Hampl V, Vanacova S, Kulda J, Flegr J. Concordance between genetic relatedness and phenotypic similarities of Trichomonas vaginalis strains. BMC Evol Biol. 2001; 1: 11.

Fraga J, Rojas L, Sariego I, Sarrı´a CA. Optimization of random amplified polymorphic DNA techniques for use in genetic studies of Trichomonas vaginalis isolates. Infect Genet Evol. 2002; 2: 73–75.

Paces J, Urbankova V, Urbanek P. Cloning and characterization of a repetitive DNA sequence specific for Trichomonas vaginalis. Mol Biochem Parasitol. 1992; 54:247 –55.

Sambrook J, Fritsch EF , Maniatis T. Molecular cloning. A laboratory manual. 2nd ed. Cold Spring Harbor, New York: Cold spring Harbor Laboratory Press, 1989.

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: 3004–3009.

Simo˜es-Barbosa A, Lobo TT, Xavier J, Carvalho SE, Leornadecz E. Trichomonas vaginalis: intrastrain polymorphisms within the ribosomal intergenic spacer do not correlate with clinical presentation. Exp Parasitol. 2005; 110: 108–113.

Crucitti T, Abdellati S, Van Dyck E, Buvé, A. Molecular typing of the actin gene of Trichomonas vaginalis isolates by PCR–restriction fragment length polymorphism. Clin Microbiol Infec. 2008; 14 (9): 844-852.

Tamura K, Peterson D, Peterson N, et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evo. 2011; 28: 2731–2739,

Laga M, Nzilla N, Goeman J. The interrelationship of sexually transmitted diseases and HIV infection implications for control of both epidemics in Africa. AIDS. 1994; 5(Suppl. 1):S55 –S63.

Kaul P, Gupta I, Sehgal R, Malla N. Trichomonas vaginalis: random amplified polymorphic DNA analysis of isolates from symptomatic and asymptomatic women in India. Parasitol Int. 2004; 53: 255–262.

Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990; 18: 6531–6535.

Arroyo R, Gonzalez-Robles A, Martinez-Palomo A, Alderette JF. Signaling of Trichomonas vaginalis for amoeboid transformation and adhesion synthesis follows cytoadherence. Mol Microbiol. 1993; 7: 299–309.

Benchimol M. Trichomonads under microscopy. Microsc Microanal. 2004; 10: 528–550.

Juliano C, Monaco G, Badeira P, Tedde G, Cappuccinelli P. Action of anticytoskeletal compounds on in vitro cytopathic effect phagocytosis, and adhesiveness of Trichomonas vaginalis. Genitour in Med. 1987; 63: 256–263.

Published
2016-12-25
How to Cite
1.
SPOTIN A, TAGHIZADEH EGHTEDAR S, SHAHBAZI A, SALEHPOUR A, SARAFRAZ S, SHARIATZADEH SA, MAHAMI-OSKOUEI M. Molecular Characterization of Trichomonas vaginalis Strains Based on Identifying Their Probable Variations in Asymptomatic Patients. IJPA. 11(4):507-14.
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