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

作者：Lehmann Lutz Eric; Hunfeld Klaus Peter^*; Emrich Thomas; Haberhausen Gerd; Wissing Heimo; Hoeft Andreas; Stueber Frank

来源：Medical Microbiology and Immunology, 2008, 197(3): 313-324.

DOI：10.1007/s00430-007-0063-0

• bacterial and fungal pathogens
• blood culture
• ICU
• multiplex PCR assay
• sepsis
• simultaneous detection

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.

• 出版日期2008-9
• 单位河北医科大学