Use of Internal Transcribed Spacer Sequence Polymorphisms as a Method for Trichomonas vaginalis Genotyping

Abstract

Objective: Trichomonas vaginalis is the most common non-viral, sexually transmitted pathogen with a worldwide distribution. The aim of the present study was to design a new genotyping tool for T. vaginalis isolates using internal transcribed spacer (ITS) sequences. Methods: First, a total of 20 cryopreserved T. vaginalis isolates were thawed and genomic DNA was isolated from fresh cultures. A polymerase chain reaction (PCR) was performed to amplify the ITS regions and the amplicons were sequenced. These sequences were aligned with others from Genbank and polymorphisms were detected. At last, each ITS sequence was given a different sequence type. Results: More than 99% homology was observed among sequences. Of 20 isolates, five had identical ITS sequence to reference (L29561) defined as ITST1. Moreover, 13 had A58 deletion (ITST10), one had C203T mutation (ITST2), and one had both A58 deletion and C203T mutation (ITST11). ITS typing of T. vaginalis sequences on Genbank revealed a total of 11 ITS types with the predominance of ITST1 (44.4%) globally. Conclusions: ITS typing seems to be an applicable and useful tool for a better understanding of molecular epidemiology as well as for the dissemination of T. vaginalis clones.

Forensic parasitology: a new frontier in criminalistics

“…However, this evidential link between a suspected rapist and victim can be further strengthened by genomic analysis of T. vaginalis in the two individuals. A wide range of hypervariable genomic markers have been used for genotyping different T. vaginalis strains, including the internal transcribed spacer [ 41 ], actin [ 42 ], tryptophanase, alanyl tRNA synthetase, and DNA mismatch repair protein genes [ 43 ]. However, more advanced techniques such as next generation multi-locus sequence typing have been used to examine dozens of polymorphic nucleotide sites to allow for a more nuanced view of the genomic diversity within T. vaginalis populations [ 44 ].…”

Trikomoniyaz tanısında farklı laboratuvar yöntemlerinin kullanılması ve Trikomoniyaz hastalarında miRNA profilinin belirlenmesi

“…Klinik örneklerden DNA izolasyonu QIAamp DNA Mini Kit'inin (Qiagen, Hilden, Almanya) kullanma talimatlarına göre yapılmıştır. PCR analizi için T. vaginalis parazitinin ITS gen bölgesi hedef alınmış ve TVITSF (5'-ACCGCCCGTCGCTCCTACCGA-3') ve TVITSR (5'-CTCCGCTTAATGAGATGCTTC-3') primerleri kullanılmıştır 8 . Amplifikasyon reaksiyonu için, 1 pmol forward primer, 1 pmol reverse primer, 1.5 U Taq DNA polymeraz, 0.5 mM dNTP, 2 mM MgCl2 ve 10 X Taq buffer (Fermentas, ABD)'dan oluşan toplam 25 µL hacimde PCR reaksiyonu hazırlanmıştır.…”

comparison of genetic diversity and phylogenetic structure of 18S gene ofTrichomonas vaginalisin Hainan and Xinjiang provinces

“…Except using wet mount microscopy, staining techniques, several methods also have used for detecting trichomoniasis, including culture, immunodiagnosis, and molecular diagnosis. [7,8,9,10].Kinds of genes or tools have used to study teh genetic properties of T.vaginalis, including actin gene, RFLP, ITS, qRT-PCR, PCR sequencing and so on [11,12,13,14] . Several researchers using microsatellites and single-copy genes acted polymorphic makers to study genetic diversity in T.vaginalis, and has been mainly identified two population [15].…”