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Sepsis pp 139-147 | Cite as

Real-Time PCR-Based Identification of Bacterial and Fungal Pathogens from Blood Samples

  • Madeleine Mai
  • Iris Müller
  • Daniela Maneg
  • Benedikt Lohr
  • Achim Haecker
  • Gerd Haberhausen
  • Klaus-Peter HunfeldEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1237)

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.

Key words

Multiplex real-time PCR Polymerase chain reaction LightCycler SeptiFast test Bloodstream infection Whole blood Rapid diagnosis Sepsis MagNA pure compact instrument 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Madeleine Mai
    • 1
  • Iris Müller
    • 1
  • Daniela Maneg
    • 1
  • Benedikt Lohr
    • 1
  • Achim Haecker
    • 2
  • Gerd Haberhausen
    • 2
  • Klaus-Peter Hunfeld
    • 1
    Email author
  1. 1.Institute of Laboratory Medicine, Microbiology and Infection ControlNorthwest Medical CentreFrankfurt/MainGermany
  2. 2.Roche Diagnostics GmbHPenzbergGermany

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