Application of Taqman RTi-PCR Assay in Specific Detection of Klebsiella pneumoniae from Surface Waters

Abstract

Klebsiella pneumoniae is an opportunistic pathogen causing nosocomial infections. The normal habitat of K. pneumoniae is the human intestines and the bacterium causes no infection in normal flora. Although, many Klebsiella infections are hospital-acquired infections, K. pneumoniae may also be transferred from environmental sources due to its widely distribution in nature. The environmental isolates of K. pneumoniae additionally pose a risk to humans as clinical isolates. The present study aims to investigate the potential of K. pneumoniae in surface waters by using PCR and RTi-PCR assays. We have optimized PCR and RTi-PCR assays with high sensitivity and specificity for K. pneumoniae. Surface waters samples were collected from different regions and analyzed by using PCR and RTi-PCR assays. The results indicated that all tested water samples are contaminated with K. pneumoniae at different levels. The RTi-PCR findings were confirmed by conventional PCR.

Keywords

• K. pneumoniae,Taqman assay,RTi-PCR,surface waters

Yüzey Sularında Bulunan Klebsiella pneumoniae'nin Özgül Olarak BelirlenmesindeTaqman RTi-PCR Yönteminin Uygulanması

Abstract

Klebsiella pneumoniae, nozokomiyal enfeksiyonlara neden olan fırsatçı bir patojendir. Klebsiella pneumoniae'nin normal habitatı insan bağırsaklarıdır ve bakteri normal florasında hiçbir enfeksiyona neden olmaz. Çoğu Klebsiella enfeksiyonu hastane kaynaklıdır ancak K. pneumoniae doğada yaygın olarak bulunmasından dolayı çevresel kaynaklardan da transfer edilebilir. Ayrıca, K. pneumoniae'nin çevresel izolatları, klinik izolatlar gibi insanlar için bir risk oluşturmaktadır. Bu çalışma ile PCR ve RTi-PCR analizlerini kullanarak yüzey sularında K. pneumoniae'nin potansiyelini araştırmak amaçlanmıştır. Klebsiella pneumoniae için yüksek hassasiyet ve özgüllük ile PCR ve RTi-PCR analizleri optimize edilmiştir. Yüzey suları örnekleri farklı bölgelerden toplanmış ve PCR ve RTi-PCR deneyleri kullanılarak analiz edilmiştir. Sonuçlar, test edilen su örneklerinin K. pneumoniae ile farklı seviyelerde kontamine olduğunu göstermiştir. RTi-PCR bulguları geleneksel PCR ile teyit edilmiştir.

Keywords

• K. pneumoniae,Taqman assay,RTi-PCR,Yüzey suları

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