Çukurova ve Şanlıurfa bölgelerinde deri lezyonlarından izole edilen leishmania sp. DNA’larının restriksiyon endonükleazlarla karşılaştırılması

Yıl 2004, Cilt: 1 Sayı: 4, 21 - 27, 01.08.2004

Fuat Dilmeç Ali Matur Soner Uzun Mehmet Karakaş Hamdi R. Memişoğlu

Öz

Amaç: Leishmaniasis, dünyanın tropikal ve subtropikal bölgelerinde yaygın olarak görülmektedir. Leishmania türlerinin tiplendirilmesinde, günümüzde klasik mikroskobik teknikler yanında DNA’ya dayalı teknikler kullanılmaktadır. Bu çalışmada, Çukurova ve Şanlıurfa bölgeleri deri lezyonlarından izole edilen Leishmania suşları arasında genotipik farklılıkların olup olmadığını saptamak amaçlanmıştır. Gereç-Yöntem: Hastaların deri lezyonu kenarlarından iğne aspirasyonu ile alınan örnekler NNN besiyerine ekilerek promastigotlar elde edilmiştir. Büyük hacimde promastigot kültürü yapıldıktan sonra promastigotlardan DNA izole edilmiş ve DNA; Eco RI, Hind III, Hae III, Hinf I, Alu I ve Rsa I restriksiyon endonükleazları ile kesilmiş ve agaroz jelinde elde edilen DNA profilleri karşılaştırılmıştır. Bulgular: 18’i Çukurova, 7’si Şanlıurfa bölgelerinden elde edilen suşlardan ve 14 referans suştan izole edilen toplam 39 DNA örneği RFLP Restriction fragment lengt polymorphisms analizi ile karşılaştırılmış ve Çukurova bölgesinden izole edilen deri Leishmania örneklerinin, Şanlıurfa bölgesi Leishmania örneklerinden daha heterojen oldukları belirlenmiştir. Sonuç: Her iki bölgeden izole edilen cutaneous leishmaniasis etkenleri arasında genotipik farklılıklar tespit edilmiştir. Tiplendirme için, PCR Polymerase Chain Reaction tekniğinin uygulanması şarttır

Anahtar Kelimeler

Jel elektroforezi, Restriksiyon endonükleaz, RFLP analizi, DNA, total DNA İzolasyonu

Comparision of leishmania sp. DNA’s isolated in the regions of Çukurova and Şanlıurfa with the restriction endonucleases

Öz

Objective: Leishmaniasis is widely seen in tropical and subtropical regions of the world. Today, DNA-based techniques are used in the identification of Leishmania sp. beside microscopic techniques. In this study, it has been aimed whether there are genotypic differences among Leishmania strains isolated from cutaneous lesions in Çukurova and Şanlıurfa regions. Materials-Methods: The samples taken by needle aspiration from the edges of skin lesion have been inoculated to change to promastigotes in NNN medium. After the promastigote culture was achieved for the large volume, DNAs from promastigotes have been isolated and they have been cut with the restriction endonucleases, such as Eco RI, Hind III, Hae III, Hinf I, Alu I, and Rsa I, and their profiles obtained on agarose gel have been compared. Results: The samples of 39 DNA isolated from 18 of Çukurova, 7 of Şanlıurfa regions, and 14 of reference strain have been compared with RFLP Restriction Fragment Length Polymorphism analysis and the cutaneous Leishmania samples isolated from Çukurova have been designated more heterogenicity than Leishmania samples taken from Şanlıurfa region. Conclusions: The genotypic differencies have been determined among the cutaneous Leishmaniasis agents isolated from either regions. PCR Polymerase Chain Reaction technique should be applied for the identification

Anahtar Kelimeler

Gel electrophoresis, Restriction endonucleases, RFLP analyses, DNA, Total DNA isolation

Kaynakça

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