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. 2017 Oct 30;10(1):537.
doi: 10.1186/s13071-017-2496-7.

Molecular typing of Trichomonas vaginalis isolates by actin gene sequence analysis and carriage of T. vaginalis viruses

Simon C Masha  1   2   3 , Piet Cools  4 , Tania Crucitti  5 , Eduard J Sanders  6 , Mario Vaneechoutte  4
Affiliations

Molecular typing of Trichomonas vaginalis isolates by actin gene sequence analysis and carriage of T. vaginalis viruses

Simon C Masha et al. Parasit Vectors. .

Abstract

Background: The protozoan parasite Trichomonas vaginalis is the most common non-viral, sexually transmitted pathogen. Although T. vaginalis is highly prevalent among women in Kenya, there is lack of data regarding genetic diversity of isolates currently in circulation in Kenya.

Methods: Typing was performed on 22 clinical isolates of T. vaginalis collected from women attending the antenatal care clinic at Kilifi County Hospital, Kenya, in 2015. Genotyping followed a previously proposed restriction fragment length polymorphism (RFLP) scheme, which involved in silico cleavage of the amplified actin gene by HindII, MseI and RsaI restriction enzymes. Phylogenetic analysis of all the sequences was performed to confirm the results obtained by RFLP-analysis and to assess the diversity within the RFLP genotypes. Additionally, we determined carriage of the four different types of Trichomonas vaginalis viruses (TVVs) by polymerase chain reaction.

Results: In silico RFLP-analysis revealed five actin genotypes; 50.0% of the isolates were of actin genotype E, 27.3% of actin genotype N, 13.6% of actin genotype G and 4.5% of actin genotypes I and P. Phylogenetic analysis was in agreement with the RFLP-analysis, with the different actin genotypes clustering together. Prevalence of TVVs was 43.5% (95% confidence interval, CI: 23.2-65.5). TVV1 was the most prevalent, present in 39.1% of the strains and 90% of the T. vaginalis isolates which harbored TVVs had more than one type of TVV. None of the isolates of actin genotype E harbored any TVV.

Conclusion: The presence of five actin genotypes in our study suggests notable diversity among T. vaginalis isolates occurring among pregnant women in Kilifi, Kenya. Isolates of the most prevalent actin genotype E lacked TVVs. We found no association between T. vaginalis genotype, carriage of TVVs and symptoms. Further studies with higher number of strains should be conducted in order to corroborate these results.

Keywords: Kenya; Kilifi; Trichomonas vaginalis; Trichomonas vaginalis viruses; Typing; actin gene.

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Conflict of interest statement

Ethics approval and consent to participate

The study was approved by the Kenya Medical Research Institute (KEMRI) Scientific and Ethics Review Unit (#3022). All participants provided written informed consent for study participation.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Phylogenetic tree for actin gene nuclotide sequences of T. vaginalis. The tree was generated using Maximum Likelihood method. Bootstrap test for 1000 replicates. Sequences from Kilifi isolates (n = 22) have accession numbers with the prefix “MF”, all other sequences were retreived from GenBank and are representative of the five actin genotypes. Scale-bar: 0.001 (1 substitution per 1000 nucleotides)
See this image and copyright information in PMC

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