Rapid identification of bacteria in blood cultures by using fluorescently labeled oligonucleotide probes.
The applicability of whole-cell hybridization for the identification of pathogenic bacteria in blood from septic patients was examined. Oligonucleotide probes, fluorescently labeled with fluorescein isothiocyanate, directed against the variable regions of the 16S rRNAs of the following bacterial species and/or genera were used: Streptococcus spp., Enterococcus faecalis, Staphylococcus aureus, coagulase-negative staphylococci (CoNS), Escherichia coli, Pseudomonas aeruginosa, and the Enterobacteriaceae family. A probe specific for the rRNAs of almost all bacteria and its complementary, reversed counterpart was used as positive and negative control, respectively. The probes were used in conjunction with a fast and simple-to-use protocol for whole-cell hybridization. This protocol yields an identification after 25 to 45 min, depending on whether the bacterium is gram positive or gram negative. A total of 182 blood samples which tested positive in a blood culture machine were investigated. All probes except for the ones for S. aureus and the CoNS showed sensitivities and specificities of 1.000. It was concluded that whole-cell hybridization is well suited for the fast screening of septic blood containing streptococci and/or enterococci or gram-negative rods.
Jansen
G J
GJ
Department of Medical Microbiology, University of Groningen, Groningen, The Netherlands.
Mooibroek
M
M
Idema
J
J
Harmsen
H J
HJ
Welling
G W
GW
Degener
J E
JE
D016470:Bacteremia / Q000175:diagnosis* / Q000382:microbiology; D001419:Bacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D001769:Blood; D003470:Culture Media; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006090:Gram-Negative Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006095:Gram-Positive Cocci / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S / Q000235:genetics
D003470:Culture Media; D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S; D016650:Fluorescein-5-isothiocyanate