Molecular typing of Trichomonas vaginalis isolates by HSP70 restriction fragment length polymorphism.

Abstract

Abstract. Subtyping isolates of Trichomonas vaginalis is an essential tool for understanding the epidemiology of this common sexually-transmitted disease. Restriction fragment length polymorphism (RFLP) analysis employing a probe from the heat-inducible cytoplasmic HSP70 gene family hybridized with EcoR I-digested genomic DNA was used in the molecular typing of Trichomonas isolates. Analysis of five American Type Culture Collection (ATCC) reference strains and 31 Jackson, Mississippi, isolates from six male and 21 female patients, revealed 10 distinct RFLP pattern subtypes of Trichomonas. The subtypes were temporally stable and cosmopolitan. The RFLP profiles seen in Maryland, Ohio, Massachusetts, and New York ATCC strains were identical to those of some Mississippi isolates, even though the samples were isolated 10-35 years apart. There was no correlation between metronidazole resistance and RFLP subtype with resistant isolates from eight patients distributed among six different subtypes.

Genetic Characterization of Trichomonas vaginalis Isolates by Use of Multilocus Sequence Typing

“…These data are in agreement with other genetic typing methods used by our laboratory and others (10,15,39,50,60). Although the average nucleotide diversity of 0.0035 differences/site is lower than some pathogenic bacteria (13), it is higher than that of its human hosts (31).…”

“…Currently, there is no "gold standard" method for genotyping isolates of T. vaginalis. A number of methods have been used in previous attempts to characterize the genetic diversity and population structure of T. vaginalis including isoenzyme analysis, repetitive sequence hybridization, random amplified polymorphic DNA, restriction fragment length polymorphism (RFLP), sequence polymorphisms in rRNA genes and intergenic regions, pulsed-field gel electrophoresis, and antigenic characterization (3,17,26,43,49,50,58,60,64,65). These methods have variable levels of reproducibility and precision in determining the genetic relatedness of isolates.…”

Human Trichomoniasis due to Trichomonas vaginalis - Current Perspectives

“…Correlation of absence of TVV and drug resistance (p<0.05) in our study supports the earlier studies (Wang and Wang, 1985;Snipes et al, 2000). However, on retrospective analysis of presence of TVV and drug sensitivity with RAPD analysis of isolates, no significant correlation was found, which is in agreement to the earlier report (Stiles et al, 2000), whereby no significant correlation between metronidazole resistance and RFLP subtypes was reported. In contrast, Vanacova et al ( 1997) have reported that all the 5 isolates refractory to metronidazole constituted single branch of tree by RAPD analysis.…”

Extensive Genetic Diversity, Unique Population Structure and Evidence of Genetic Exchange in the Sexually Transmitted Parasite Trichomonas vaginalis

“…Crude genotyping markers such as random amplified polymorphic DNA (RAPDs) and restriction fragment length polymorphisms (RFLPs), have indicated genetic variation among T. vaginalis isolates and have inconclusively detected evidence of population structure [19]–[25]. These methods, however, are highly sensitive to contaminating DNA or to slight variation in conditions, which may influence the interpretation of data collected with these techniques.…”