Medical Microbiology and Immunology

, Volume 197, Issue 3, pp 313–324

A multiplex real-time PCR assay for rapid detection and differentiation of 25 bacterial and fungal pathogens from whole blood samples

  • Lutz Eric Lehmann
  • Klaus-Peter Hunfeld
  • Thomas Emrich
  • Gerd Haberhausen
  • Heimo Wissing
  • Andreas Hoeft
  • Frank Stüber
Original Investigation

Abstract

Early detection of bloodstream infections (BSI) is crucial in the clinical setting. Blood culture remains the gold standard for diagnosing BSI. Molecular diagnostic tools can contribute to a more rapid diagnosis in septic patients. Here, a multiplex real-time PCR-based assay for rapid detection of 25 clinically important pathogens directly from whole blood in <6 h is presented. Minimal analytical sensitivity was determined by hit rate analysis from 20 independent experiments. At a concentration of 3 CFU/ml a hit rate of 50% was obtained for E. aerogenes and 100% for S. marcescens, E. coli, P. mirabilis, P. aeruginosa, and A. fumigatus. The hit rate for C. glabrata was 75% at 30 CFU/ml. Comparing PCR identification results with conventional microbiology for 1,548 clinical isolates yielded an overall specificity of 98.8%. The analytical specificity in 102 healthy blood donors was 100%. Although further evaluation is warranted, our assay holds promise for more rapid pathogen identification in clinical sepsis.

Keywords

Bacterial and fungal pathogens Blood culture ICU Multiplex PCR assay Sepsis Simultaneous detection 

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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Lutz Eric Lehmann
    • 1
  • Klaus-Peter Hunfeld
    • 3
  • Thomas Emrich
    • 4
  • Gerd Haberhausen
    • 4
  • Heimo Wissing
    • 2
  • Andreas Hoeft
    • 1
  • Frank Stüber
    • 1
  1. 1.Klinik und Poliklinik für Anästhesie und operative IntensivmedizinUniversitätsklinikum BonnBonnGermany
  2. 2.Zentrum der Anästhesiologie und WiederbelebungUniversitätsklinikum Frankfurt am MainFrankfurt am MainGermany
  3. 3.Institut für Medizinische Mikrobiologie und KrankenhaushygieneUniversitätsklinikum Frankfurt am MainFrankfurt am MainGermany
  4. 4.Roche Diagnostics GmbHPenzbergGermany

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