Rapid Detection of Klebsiella pneumoniae from Blood Culture Bottles by Real-Time PCR
Prathiba Kurupati, Carol Chow, Gamini Kumarasinghe, Chit Laa Poh
Abstract
Prathiba Kurupati, Carol Chow, Gamini Kumarasinghe, Chit Laa Poh
Abstract
A LightCycler real-time PCR hybridization probe-based assay which detects a partial Klebsiella pneumoniae 16S rRNA gene was developed for the rapid identification of K. pneumoniae directly from growth-positive blood culture bottles (BACTEC 9240 system) within 2 h. No cross-reactivity was observed with 65 negative-control blood cultures that grew bacteria other than K. pneumoniae and 48 negative blood cultures from double-blind experiments, thus demonstrating 100% specificity when compared to results of conventional biochemical characterization. The assay also showed 100% sensitivity, as it correctly identified all 142 positive-control blood cultures and 4 from double-blind trials.
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A LightCycler real-time PCR hybridization probe-based assay which detects a partial Klebsiella pneumoniae 16S rRNA gene was developed for the rapid identification of K. pneumoniae directly from growth-positive blood culture bottles (BACTEC 9240 system) within 2 h. No cross-reactivity was observed with 65 negative-control blood cultures that grew bacteria other than K. pneumoniae and 48 negative blood cultures from double-blind experiments, thus demonstrating 100% specificity when compared to results of conventional biochemical characterization. The assay also showed 100% sensitivity, as it correctly identified all 142 positive-control blood cultures and 4 from double-blind trials.
Key concepts: Klebsiella pneumoniae, Microbiology, Blood culture, Bacteria, 16S ribosomal RNA, Biology, Enterobacteriaceae, Klebsiella