Use of a multiplex-polymerase chain reaction for detection of Salmonella and Chlamydophila psittaci from caged birds

Abstract

In this study, development of a multiplex-PCR (m-PCR) assay for identification of Salmonellosis and Psittacosis in caged birds was aimed. DNAs extracted from Salmonella spp. strains from the collection of our department and Salmonella spp. isolates from caged birds in addition to the extracted DNA samples belong to two different Chlamydophila psittaci clinical strains (strain 6BC and a strain isolated from a parakeet), constituted the material of the study. A m-PCR assay was developed which use iroBF and iroBR primers to specifically amplify 606 bp sequence of iroB gene of Salmonella spp. DNA in the same reaction mix and with the same conditions, CpsiA and CpsiB primers were used to amplify 300 bp sequence of pmp gene of C. psittaci. The specificity and the sensitivity of the assay were determined by using positive controls (Salmonella and Chlamydia DNA samples). We believe that the newly developed m-PCR assay is a fast, reliable and an economic assay, which has the potential for direct identification of Salmonella spp. and C. psittaci from the clinical samples

Keywords

• Caged birds
• Chlamydophila psittaci
• identification
• multiplex-PCR
• Salmonella spp

Kafes kuşlarında Salmonella ve Chlamydophila psittaci’nin saptanmasında multiplex-polimeraz zincirreaksiyonu tekniğinin kullanılması

Abstract

Bu çalışmada, Salmonellosis ve Psittacosis’in tanısında kullanılmak üzere bir multipleks-polimeraz zincir reaksiyonu (m-PZR) tekniği geliştirilmesi amaçlandı. Ankara Üniversitesi Veteriner Fakültesi Mikrobiyoloji Anabilim Dalı suş koleksiyonunda yer alan Salmonella suşları, kafes kuşu orijinli Salmonella izolatları ve bunlardan izole edilen DNA örnekleri dışında yurt dışından sağlanan ve iki farklı Chlamydophila psittaci klinik suşundan izole edilmiş DNA örnekleri çalışma materyalini oluşturdu. Çalışmada, Salmonella türlerinde iroB genini kodlayan sekansın 606 bp’lik porsiyonunu spesifik olarak amplifiye eden iroBF ve iroBR primer çifti ile Chlamydophila psittaci’nin pmp genlerinin 300 bp’lik sekansını amplifiye eden CpsiA ve CpsiB primer çiftinin aynı reaksiyon karışımı ve şartlarında çalışacağı bir m-PZR tekniği geliştirildi. Pozitif kontrol Chlamydophila psittaci DNA’larının dışında, kendi laboratuvar izolatları ve suşlarından izole edilen DNA örnekleri kullanılarak tekniğin spesifite ve sensitivitesi de belirlendi. Sonuç olarak, klinik materyallerden direkt tanı potansiyeli de bulunan tekniğin kafes kuşlarında Salmonella spp. ve C. psittaci’nin tanısında hızlı, güvenilir ve ekonomik bir teknik olduğu düşünülmektedir

Keywords

• Chlamydophila psittaci
• kafes kuşları
• multiplex-PCR
• Salmonella spp.
• tanı

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