Detection and Molecular Characterization of Canine Coronavirus Based on Partial Membrane Gene Sequences

Öz

Canine coronavirus (CCoV) infection in dogs is common all over the world and progresses with gastroenteritis findings. Infection as a result of complications with secondary factors may result in death, especially in puppies. The virus, which is excreted in the feces, spreads indirectly through the contamination of food, water, and the environment. This study, it was aimed at revealing the CCoV infection and obtaining current molecular information about the infection. In addition, molecular characterization of CCoV strains circulating in the region was made based on the M (membrane protein) gene. The study material consisted of stool samples from 12 dogs with gastroenteritis findings. The amplified PCR products were subjected to sequence analysis and a phylogenetic tree was constructed by comparing them with different reference CCoV isolates from GenBank. In the phylogenetic tree, 1 of the 5 positive samples was determined to be CCoV-I, and 4 samples were determined to be CCoV-IIa. It was determined that the strains obtained were 85.4 - 97.7% similar among themselves and 82.7-98% similar to other strains obtained from GenBank. As a result of study, current molecular information about CCoV circulating in the Balıkesir region was obtained. With this study, it is thought that new research on the existence and molecular epidemiology of CCoV infection in Türkiye will make important contributions to vaccine studies and the control of infection.

Anahtar Kelimeler

• Canine coronavirus
• PCR
• phylogenetic analysis.

Canine Coronavirusun Tespiti ve Kısmi Membran Gen Dizisi Temelli Moleküler Karakterizasyonu

Öz

Köpeklerin Koronavirus (CCoV) enfeksiyonu tüm dünyada yaygın olarak görülmekte ve gastroenteritis bulguları ile seyretmektedir. Sekonder etkenlerle komplikasyon sonucunda enfeksiyon, özellikle yavru köpeklerde ölümle sonuçlanabilir. Dışkı ile atılan virus, dışkının yeme, suya ve çevreye bulaşmasıyla indirekt olarak yayılır. Yapılan bu çalışmada CCoV enfeksiyonunun ortaya konması, enfeksiyona ilişkin güncel moleküler bilgilerin elde edilmesi amaçlanmıştır. Ayrıca, bölgede dolaşan CCoV suşlarının M (membran proteini) genine dayalı moleküler karakterizasyonu da yapılmıştır. Çalışma materyalini, gastroenteritis semptomları gösteren 12 adet köpeğe ait gaita örneği oluşturdu. PCR ürünleri saflaştırdıktan sonra sekans analizine tabi tutuldu ve GenBank veri tabanından sağlanan farklı referans CCoV izolatları ile karşılaştırılarak filogenetik ağaç oluşturuldu. Filogenetik analiz sonucu 5 pozitif örnekten, 1 adedinin CCoV-I, 4 adet örneğin ise CCoV-IIa olduğu tespit edildi. Elde edilen suşların kendi aralarında %85.4-%97.7 oranında, GenBank veri tabanından elde edilen diğer suşlarla aralarında %82,7-%98 oranında benzerlik olduğu tespit edildi. Çalışma sonucunda, Balıkesir bölgesinde sirküle olan CcoV’ye yönelik güncel moleküler bilgiler elde edilmiştir. Bu çalışma ile, Türkiye’de CCoV enfeksiyonunun varlığına ve moleküler epidemiyolojisine ilişkin yapılacak yeni araştırmaların aşı çalışmalarına ve enfeksiyonun kontrolüne önemli katkılar sağlayacağı düşünülmektedir.

Anahtar Kelimeler

• Köpek koronavirus
• filogenetik analiz
• PZR.

Kaynakça

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