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2020, Cilt 50, Sayı 2, Sayfa(lar) 086-094
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Designing of the Primers and Probes for Real Time Polymerase Chain Reaction of Cytochrome B Gene Region in Leishmaniasis: A Pilot Study
Ahmet Özbilgin1, Bakiye Göker Bağca2, Mehmet Harman3, İbrahim Çavuş1, Tuğba Kaya4, Ahmet Yıldırım1, Cumhur Gündüz2
1Manisa Celal Bayar Üniversitesi Tıp Fakültesi, Tıbbi Parazitoloji Anabilim Dalı, Manisa, Türkiye
2Ege Üniversitesi Tıp Fakültesi, Tıbbi Biyoloji Anabilim Dalı, İzmir, Türkiye
3Dicle Üniversitesi Tıp Fakültesi, Dermatoloji Anabilim Dalı, Diyarbakır, Türkiye
4Mustafa Kemal Üniversitesi Tıp Fakültesi, Parazitoloji Anabilim Dalı, Hatay, Türkiye
Keywords: Cytochrome B, RT-PCR, Leishmania

Objective: In this study, it was aimed to design primers and probes to identify Leishmania species (Leishmania donovani, Leishmania major, Leishmania tropica and Leishmania infantum) through Real Time Polymerase Chain Reaction using the cytochrome b gene region.

Method: The Leishmania strains, which have been cryopreserved and stored in liquid nitrogen in the Parasite Bank of Manisa Celal Bayar University, were transferred to culture media after being revived under the appropriate conditions and their DNA isolations were conducted using a commercial kit. Real Time Polymerase Chain Reaction melting analyses were conducted for the extracted DNA by utilizing both the candidate primers and probes of cytochrome b gene region, newly designed for Leishmania, and the primers and probes, designed for internal transcribed spacer-1 gene region we had previously used for control.

Results: Leishmania tropica, Leishmania major, Leishmania infantum, Leishmania donovani, Leishmania aethiopica and Leishmania infantum/donovani hybrid species were managed to be genotyped succesfully using the primers and probes of the newly designed cytochrome b gene region.

Conclusion: It has been observed that Old World Leishmania species (Leishmania tropica, Leishmania major, Leishmania infantum, Leishmania donovani, Leishmania aethiopica and Leishmania infantum/donovani hybrid), rather than New World Leishmania species (Leishmania brazilensis and Leishmania amazonensis), were successfully genotyped by the primers and probes of the newly designed cytochrome b gene region.


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