Real-time PCR for detection and differentiation of gram-positive and gram-negative bacteria

J Clin Microbiol. 2002 Nov;40(11):4304-7. doi: 10.1128/JCM.40.11.4304-4307.2002.

Abstract

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.

Publication types

  • Evaluation Study

MeSH terms

  • Bacterial Infections / microbiology*
  • Bacterial Typing Techniques
  • Body Fluids / microbiology*
  • DNA Probes
  • DNA, Ribosomal / analysis
  • Fluorescent Dyes
  • Gentian Violet
  • Gram-Negative Bacteria / classification*
  • Gram-Negative Bacteria / genetics
  • Gram-Negative Bacteria / isolation & purification
  • Gram-Positive Bacteria / classification*
  • Gram-Positive Bacteria / genetics
  • Gram-Positive Bacteria / isolation & purification
  • Humans
  • Intensive Care Units
  • Phenazines
  • Polymerase Chain Reaction / methods*
  • RNA, Ribosomal, 16S / genetics

Substances

  • DNA Probes
  • DNA, Ribosomal
  • Fluorescent Dyes
  • Gram's stain
  • Phenazines
  • RNA, Ribosomal, 16S
  • Gentian Violet