Real-time PCR-based identification of bacterial and fungal pathogens from blood samples

Methods Mol Biol. 2015:1237:139-47. doi: 10.1007/978-1-4939-1776-1_14.

Abstract

Latest major contributions in the field of sepsis diagnostics result from advances in PCR technologies permitting new standards in speed and quality, given the fact that a timely diagnosis is the decisive factor to the survival of patients with bloodstream infections.Multiplex real-time PCR is a quantitative method for simultaneous amplification and detection of different targeted DNA molecules within hours. Nevertheless, various studies have shown a number of technical shortcomings as well as a high heterogeneity in sensitivity.The present method allows the standardized and rapid detection and identification of 25 common bacteria and fungi responsible for bloodstream infections from whole blood samples by using LightCycler(®) SeptiFast (LC-SF) test, based on real-time PCR.

MeSH terms

  • Automation, Laboratory
  • Bacteremia / diagnosis*
  • Bacteremia / microbiology
  • Bacteremia / pathology
  • Bacteria / genetics
  • Bacteria / growth & development
  • Bacteria / isolation & purification
  • Blood Specimen Collection
  • Culture Media / chemistry
  • DNA, Bacterial / genetics
  • DNA, Bacterial / isolation & purification*
  • DNA, Fungal / genetics
  • DNA, Fungal / isolation & purification*
  • Fungemia / diagnosis*
  • Fungemia / microbiology
  • Fungemia / pathology
  • Fungi / genetics
  • Fungi / growth & development
  • Fungi / isolation & purification
  • Humans
  • Molecular Diagnostic Techniques
  • Multiplex Polymerase Chain Reaction / methods
  • Multiplex Polymerase Chain Reaction / standards*
  • Real-Time Polymerase Chain Reaction / methods
  • Real-Time Polymerase Chain Reaction / standards*
  • Time Factors

Substances

  • Culture Media
  • DNA, Bacterial
  • DNA, Fungal