İnci Başak MÜŞTAK, SEYYİDE SARIÇAM, Hamit Kaan MÜŞTAK

Comparison of Internal Transcribed Spacer Region Sequencing and Conventional Methods Used in the Identification of Fungi Isolated from Domestic Animals

The aim of this study was to compare gold standard conventional culture method and internal transcribed spacer (ITS) sequence-based analysis in the identification of fungi isolated from domestic animals. A total of 35 animals (15 cats, 11 dogs, 4 horses and 5 chickens) suspected for fungal infection were examined. Of the 35 samples, 20 were found to be positive for fungal culture. Among positive samples 8 (40%) were predominantly found to be dermatophyte species by conventional methods. The ITS regions (ITS1, ITS2 and complete ITS) of fungal isolates were also amplified, sequenced and the results were compared with conventional culture method. The identification results of 18 (90%) fungal species were found to be compatible with both conventional culture and sequencing methods. Comparison of the results demonstrated that, complete ITS regions gene sequencing could be used for the identification of medically important fungi rapidly. Since the results of complete ITS regions gene sequencing were found to be compatible with the results of phenotypic identification, it can be concluded that ITS regions gene sequencing of fungal isolates can be also used as a confirmative tool of conventional culture methods.

Evcil Hayvanlardan İzole Edilen Mantarların Teşhisinde Kullanılan Konvansiyonel ve ITS Dizi Analizi Metotlarının Karşılaştırılması

Bu çalışmanın amacı, evcil hayvanlardan izole edilen mantar türlerinin identifikasyonunda altın standart olan konvansiyonel kültür yöntemi ve Internal Ara Bölgeler (ITS) dizi analizi yönteminin karşılaştırılmasıdır. Çalışmada, mantar enfeksiyonlarından şüphelenilen toplam 35 hayvan (15 kedi, 11 köpek, 4 at ve 5 tavuk) incelendi. 35 örnek içinde 20 tanesinin mantar kültürleri pozitif bulundu. Pozitif örnekler arasında 8 (%40) tanesi konvansiyonel yöntemlerle ağırlıklı olarak dermatofit türleri olarak bulundu. İzolatların ITS bölgeleri (ITS1, ITS2 ve ITS bölgesinin tamamı) amplifiye edildi, sekanslandı ve elde edilen sonuçlar kültür yöntemiyle karşılaştırıldı. Hem konvansiyonel hem de dizi analizi yöntemiyle elde edilen identifikasyon sonuçları 18 (%90) örnekte birbiri ile uyumlu bulundu. Bu sonuçlar doğrultusunda, ITS bölgesinin tamamının dizi analizi, mantarların hızlı teşhisi için kullanılabileceğini ortaya koydu. ITS gen bölgesinin tamamına ait dizi analizi sonuçlarının fenotipik identifikasyon sonuçları ile uyumlu olması, ITS gen bölgesi dizi analizinin konvansiyonel yöntemlerin doğrulanmasında bir araç olarak kullanılabileceğini ortaya koymuştur.

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