A Real-Time ITS1-PCR Based Method in the Diagnosis and Species Identification of Leishmania Parasite from Human and Dog Clinical Samples in Turkey

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dc.authoridTetik Vardarli, Asli/0000-0001-9890-3256
dc.authoridGunduz, Cumhur/0000-0002-6593-3237
dc.contributor.authorToz, Seray Ozensoy
dc.contributor.authorCulha, Gulnaz
dc.contributor.authorZeyrek, Fadile Yildiz
dc.contributor.authorErtabaklar, Hatice
dc.contributor.authorAlkan, M. Ziya
dc.contributor.authorVardarli, Asli Tetik
dc.contributor.authorGunduz, Cumhur
dc.date.accessioned2024-09-18T21:06:33Z
dc.date.available2024-09-18T21:06:33Z
dc.date.issued2013
dc.departmentHatay Mustafa Kemal Üniversitesien_US
dc.description.abstractHuman visceral leishmaniasis (VL) caused by L. infantum and cutaneous leishmaniasis (CL) caused by L. tropica and L. infantum have been reported in Turkey. L. infantum is also responsible for canine leishmaniasis (CanL) and it is widely common in the country. The main aim of the present study was to design a real-time PCR method based on the internal transcribed spacer 1 (ITS1) region in the diagnosis of all clinical forms of leishmaniasis in Mediterranean, and to identify the species directly from clinical samples. Totally, 315 clinical specimens, human/canine visceral (blood, bone marrow, lymph node) and cutaneous (lesion aspiration) samples, and 51 Turkish Leishmania isolates typed by isoenzymatic method were included in the study. For optimization, DNA samples of the 34 strains were amplified by conventional ITS1-PCR and then sequenced for designing the primers and probes, allowing the species identification. Following the validation with the isolates, the test was applied on clinical samples and melting temperatures were used for genotyping. A group of PCR products were further sequenced for confirmation and assigning the inter-and intraspecies heterogeneity. The diagnosis of leishmaniasis is successfully achieved by the new real-time PCR method, and the test identified 80.43% of human and canine VL samples as L. infantum and 6.52% as L. tropica; 52.46% of CL samples as L. infantum and 26.90% as L. tropica. In 13.04% of visceral and 20.62% of cutaneous samples, two peaks were observed. Hovewer, the higher peak was found to be concordant with the sequencing results in 96.96%, in terms of species identification. The real-time ITS1 PCR assay clearly identified the leishmanial species in 81.58% of all clinical samples. Genotypic variations of Leishmania parasites in Turkey within species and intraspecies were observed, and L. tropica is also found as causative agent of human and canine VL in Turkey.en_US
dc.description.sponsorshipScientific and Techological Research Council of Turkey [107S154]en_US
dc.description.sponsorshipThis study is financially supported by the Scientific and Techological Research Council of Turkey (Project No: 107S154). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.en_US
dc.identifier.doi10.1371/journal.pntd.0002205
dc.identifier.issn1935-2735
dc.identifier.issue5en_US
dc.identifier.pmid23675543en_US
dc.identifier.scopus2-s2.0-84878487990en_US
dc.identifier.scopusqualityQ1en_US
dc.identifier.urihttps://doi.org/10.1371/journal.pntd.0002205
dc.identifier.urihttps://hdl.handle.net/20.500.12483/13691
dc.identifier.volume7en_US
dc.identifier.wosWOS:000319994400012en_US
dc.identifier.wosqualityQ1en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherPublic Library Scienceen_US
dc.relation.ispartofPlos Neglected Tropical Diseasesen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectFragment Length Polymorphismen_US
dc.subjectWorld Cutaneous Leishmaniasisen_US
dc.subjectCanine Visceral Leishmaniasisen_US
dc.subjectOld-Worlden_US
dc.subjectGenetic-Heterogeneityen_US
dc.subjectTropicaen_US
dc.subjectEpidemiologyen_US
dc.subjectInfantumen_US
dc.subjectDonovanien_US
dc.subjectStrainsen_US
dc.titleA Real-Time ITS1-PCR Based Method in the Diagnosis and Species Identification of Leishmania Parasite from Human and Dog Clinical Samples in Turkeyen_US
dc.typeArticleen_US

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