Real-time PCR for detection and differentiation of gram-positive and gram-negative bacteria.
We developed a consensus real-time PCR protocol that enables us to detect spiked bacterial 16S DNA from specimens such as water, urine, plasma, and sputum. The technique allows an exact Gram stain classification of 17 intensive care unit-relevant bacteria by means of fluorescence hybridization probes. All tested bacteria were identified correctly, and none gave a false-positive signal with the opposite Gram probe.
Klaschik
Sven
S
Department of Anesthesiology and Intensive Care Medicine, University of Bonn, Bonn, Germany. klaschik@ukb.uni-bonn.de
Lehmann
Lutz E
LE
Raadts
Ansgar
A
Book
Malte
M
Hoeft
Andreas
A
Stuber
Frank
F
D001424:Bacterial Infections / Q000382:microbiology*; D015373:Bacterial Typing Techniques; D001826:Body Fluids / Q000382:microbiology*; D015342:DNA Probes; D004275:DNA, Ribosomal / Q000032:analysis; D005456:Fluorescent Dyes; D005840:Gentian Violet; D006090:Gram-Negative Bacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D006094:Gram-Positive Bacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D007362:Intensive Care Units; D010619:Phenazines; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics
D015342:DNA Probes; D004275:DNA, Ribosomal; D005456:Fluorescent Dyes; C035863:Gram's stain; D010619:Phenazines; D012336:RNA, Ribosomal, 16S; D005840:Gentian Violet