Quantitation of Pseudomonas aeruginosa in wound biopsy samples from bacterial culture to rapid `real-time polymerase chain reaction 英文参考文献.docVIP

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Quantitation of Pseudomonas aeruginosa in wound biopsy samples from bacterial culture to rapid `real-time polymerase chain reaction 英文参考文献.doc

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Quantitation of Pseudomonas aeruginosa in wound biopsy samples from bacterial culture to rapid `real-time polymerase chain reaction 英文参考文献

/content/4/4/255 Primary research Quantitation of Pseudomonas aeruginosa in wound biopsy samples: from bacterial culture to rapid ‘real-time’ polymerase chain reaction Jean-Paul Pirnay*?, Daniel De Vos??, Luc Duinslaeger*, Pascal Reper*, Christian Vandenvelde*, Pierre Cornelis? and Alain Vanderkelen* *Queen Astrid Military Hospital, Neder-Over-Heembeek, ?Flanders Interuniversity Institute of Biotechnology, Sint-Genesius-Rode, and ?Innogenetics, Neder-Over-Heembeek, Belgium Received: 9 January 2000 Crit Care 2000, 4:255–261 Revisions requested: 29 February 2000 Revisions received: 8 June 2000 Accepted: 14 June 2000 The electronic version of this article can be found online at /content/4/4/255 Published: 7 July 2000 ? Current Science Ltd Statement of findings 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 1 h. 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 Synopsis Introduction: Early diagnosis of wound colonisation or sample was taken from each dilution tube as a template for the three PCR-based methods. Serial P aeruginosa dilutions (see above) were added to uninfected cadaveric skin. The reconstituted biopsy samples were homogenized using a tissue tearer and DNA was prediction of wound sepsis provides an opportunity for therapeutic intervention. There is need for qualitative and quantitative tests that are more rapid than bacterial culture. Pseudomonas aerugino