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Quantitation of Pseudomonas aeruginosa in wound biopsy samples: from bacterial culture to rapid `real-time' polymerase chain reaction

Jean-Paul Pirnay1,2, Daniel De Vos3,2, Luc Duinslaeger1, Pascal Reper1, Christian Vandenvelde1, Pierre Cornelis2 and Alain Vanderkelen1

Author Affiliations

1 Queen Astrid Military Hospital, Neder-Over-Heembeek, Belgium

2 Flanders Interuniversity Institute of Biotechnology, Sint-Genesius-Rode, Belgium

3 Innogenetics, Neder-Over-Heembeek, Belgium

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Critical Care 2000, 4:255-262 doi:10.1186/cc702

Published: 7 July 2000

Abstract

We developed a real-time detection (RTD) polymerase chain reaction (PCR) with rapid thermal cycling to detect and quantify Pseudomonas aeruginosa in wound biopsy samples. This method produced a linear quantitative detection range of 7 logs, with a lower detection limit of 103 colony-forming units (CFU)/g tissue or a few copies per reaction. The time from sample collection to result was less than 1h. RTD-PCR has potential for rapid quantitative detection of pathogens in critical care patients, enabling early and individualized treatment.

Keywords:
burn wound; polymerase chain reaction; Pseudomonas aeruginosa; quantitation; sepsis