Differential diagnosis of Fasciola hepatica from different animal hosts by PCR

Amaç: Bu çalışmanın amacı koyun ve keçiden elde edilen Fasciola hepatica'nın PCR ile ayırıcı teşhisinin yapılmasıdır.Gereç ve Yöntem: Örnek materyal (F. hepatica) koyun ve keçilerden elde edildi. Elde edilen parazitler birkaç kez pH:7.4 fosfat bufer solusyonunda yıkandıktan sonra, genomik DNA ekstraksiyonu amacıyla fenol ekstraksiyon/etanol presipitasyon metodu uygulandı. PCR koşullarının optimizasyonunda her bir F. hepatica'nın DNA'sını amplifiye edebilmek için rastgele 5 adet primer seçildi.Bulgular: Hem koyun, hemde keçilerden elde edilen F. hepatica'ların RAPD-PCR tekniği ile amplifikasyonu sonucu farklı büyüklük ve sayıda bandlar elde edildi. Elde edilen bu bandların büyüklükleri ve sayıları sayesinde F. hepatica'ya ait genetik varyasyonlar identifiye edildi. Koyun ve keçi orjinli parazitlere (F. hepatica) ait bu bandlar tür spesifiktir.Öneri: Bu çalışma ile koyun ve keçilerde bulunan F. hepatica'nın ayırıcı teşhisinde RAPD-PCR tekniğinin kullanılmasının daha uygun olacağı saptanmıştır.

Aim: The objective of this study was evaluated differentiational diagnosis of Fasciola hepatica from sheep and goat by PCR.Materials and Methods: F. hepatica were obtained from sheep and goat. These parasites were washed several times with pH:7.4 phosphate-buffered saline and phenol extraction/ ethanol precipitation method was employed in genomic DNA extraction. In order to optimization of the PCR conditions, five primers were selected to amplify the DNA of each F. hepatica.Results: Both sheep and goat were amplified products in different numbers and sizes by RAPD-PCR technique. The results show that different primers gave different bands (fragments) and allowed to identify genetic variations of F. hepatica. These bands were species-specific to F. hepatica from goat and sheep.Conclusions: The RAPD-PCR method can be useful for the differentiational diagnosis of F. hepatica of sheep and goat.

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