İstanbul Yöresinde Sağlıklı ve Klinik Belirti Gösteren Köpeklerde Helicobacter pylori Varlığının Kültür, Polimerase Chain Reaction (PCR) ve Restriction Fragment Length Polymorphism (RFLP) ile Saptanması

Günümüzde Helicobacter pylori’nin dünya nüfusunun neredeyse yarısını infekte eden bir patojen olması ve gün geçtikçe yayılması, kedi ve köpeklerin insanlar için infeksiyon kaynağı olabileceğini düşündürmekte, bu konuda araştırmalara yönlenmeye gereksinim oluşturmaktadır. Bu araştırmada, köpeklerde de insanlardakine benzer infeksiyona neden olabileceği düşünülen H. pylori’nin varlığı köpek dışkı örneklerinde araştırılmıştır. Bu amaçla İstanbul’da bulunan toplam 96 adet pet ve sokak köpeğine ait dışkı örnekleri toplanmıştır. Dışkı örnekleri Polymerase Chain Reaction (PCR) ile incelenmiş, Helicobacter DNA’sı saptanan örneklerde H. pylori’de bulunan CagA geninin saptanması amacıyla Restriction Fragmenth Length Polymorphism (RFLP) ile H.pylori spesifik bandlar araştırılmıştır. Ayrıca çalışmada PCR-RFLP ile H. pylori saptanan dışkı örneklerinden bakteriyolojik kültür yapılarak moleküler yöntemler ile kültür tekniği de karşılaştırılmıştır. Helicobacter cins spesifik primerler kullanarak yapılan PCR uygulaması sonucu 96 örneğin 65 (% 67,7) inden elde edilen 375 bp band sonuçlarını takiben HinfI enzim kesimi ile 12 (% 18,46) DNA örneğinde H.pylori spesifik bandlar (196 bp, 119 bp, 60 bp) saptanmıştır. H. pylori DNA’sı saptanan 12 köpeğin dışkı örneğinden yapılan bakteriyolojik inceleme sonucunda ise örneklerin hiçbirinden H. pylori izole edilememiştir.

Detection Of Helicobacter Pylori In Healthy Dogs And Dogs With The Clinical Semptoms By Culture, Polimerase Chain Reaction ( PCR ) And Restriction Fragment Length Polymorphism ( RFLP ) In İstanbul Province

Helicobacter pylori is one of the most widespread infections in human beings. It is estimated that approximately half of the human population is infected with this pathogen. It has been considered that dogs and cats may serve as a source for human infection, and therefore, further researches on this subject are needed. In this study, precence of H.pylori which is considered as a source of similar infection in dogs, was investigated in dogs’ stool speciments. For this purpose, stool speciments were collected from 96 pet and sheltered dogs in Istanbul. Stool speciments were investigated by PCR and samples which contain Helicobacter DNA were objected to Restriction Fragment Lenght Polymorphism (FRLP) analyses to detect bands of H.pylori specific CagA gen. Also, samples found to be positive by PCR-RFLP were examinated bacteriologically for comparison of culture and moleculer methods. As a result of PCR investigation with Helicobacter genus specific primers, 375 bp bands were obtained for 65 (67.7 %) of the speciments. After that H.pylori specific bands (196 bp, 119 bp, 60 bp) were detected from 12 (18.46 %) of DNA samples by HinfI enzim restriction. H.pylori was not isolated from fecal speciments of 12 H.pylori DNA positive dogs.

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