Rapid detection of Klebsiella pneumoniae from blood culture bottles by real-time PCR.
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.
Kurupati
Prathiba
P
Department of Microbiology, National University of Singapore, Singapore 117597, Republic of Singapore.
Chow
Carol
C
Kumarasinghe
Gamini
G
Poh
Chit Laa
CL
D001769:Blood; D003199:Computer Systems; D003470:Culture Media; D017931:DNA Primers; D004275:DNA, Ribosomal / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D007711:Klebsiella pneumoniae / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
D003470:Culture Media; D017931:DNA Primers; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S