Türkiyenin Çukurova Bölgesinde Kliniksel Materyalde Bulunan Leishmania Parazitlerinin Polimeraz Zincir Reaksiyonu Ile Araştırılması

Year 2008, Volume: 5 Issue: 1, 10 - 14, 01.04.2008

Fuat Dılmec Ali Matur Emre Alhan Soner Uzun Mehmet Karakas Derya Alabaz Nurten Aksoy Feridun Akkafa Necmi Aksaray Hamdi R

Abstract

Giriş: Leishmania cinsi parazitlerinin sebep olduğu Leishmaniazis, oldukça önemli bir sağlık problemi olduğu düşünülür. Klinik materyalde Leishmania parazitlerinin belirlenmesi ve tür ayırımında polimeraz zincir reaksiyonurestriksiyon fragman uzunluğu polimorfizm metodunu PCR-RFLP kullanmayı ve mikroskobik inceleme sonuçlarıyla karşılaştırmayı amaçladık. Metotlar: Kırk beş deri lezyonu, 9 kemik iliği ve 4 referans türünden oluşan toplam 58 örnek, bu çalışmaya alındı. Örnekler hem DNA elde edilmesi ve hem de preparat hazırlanmasında kullanıldı. DNA’lar, Leishmania alt-cinsinin belirlenmesi için PCR yöntemi ile çoğaltıldı ve PCR ürünü, tür ayırımı için HaeIII ile kesildi. Sonuçlar: Visseral Leishmania parazitleri Leishmania infantum olarak belirlendi. Deri leişmanyazın 45 hastanın, çoğunda Leishmania tropica belirlenirken, diğerleri Leishmania donovani’ye benzer bulundu. Aynı zamanda PCR’ın pozitiflik oranı %100 , mikroskobik inceleme %67 sonuçlarından daha fazla olduğu saptandı. Çıkarım: PCR-RFLP tekniği, Leishmania türlerinin tespit ve ayırımında çok hassas olduğu gözlendi

Keywords

Tipleme, Leishmaniazis, PCR, RFLP

Investigation Of Leishmania Parasites From Clinical Samples Using Polymerase Chain Reaction Technique In Cukurova Region, Turkey

Abstract

Background: Leishmaniasis, caused by parasites of the genus Leishmania is still considered an important health problem. We aimed to appraise a Polymerase Chain Reaction-Restriction Fragment Length Polymorphism PCR-RFLP method for the detection and species differentiation of Leishmania parasites in clinical samples; and to compare with the results of microscopic examination. Methods: Totally 58 samples were taken from 45 skin lesion, nine bone marrow aspirates, and four reference strains. The samples were used for both DNA and smear-slide preparations. The DNAs were amplified by PCR for the detection of Leishmania subgenus and PCR products were restricted with HaeIII for the species differentiation. Results: The visceral Leishmania parasites VL were genotyped as Leishmania infantum. Of 45 patients with CL, most of them were analyzed as Leishmania tropica, but the others were characterized similar to Leishmania donovani reference. It was also determined positivity rate in PCR 100% was higher than microscopic examination about 67% . Conclusions: PCR-RFLP technique appears to be most sensitive for the detection and the differentiation of Leishmania species

Keywords

Identification, Leishmaniasis, PCR, RFLP.

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