Quantitation ofPseudomonas aeruginosain wound biopsy samples: from bacterial culture to rapid `real-time' polymerase chain reaction

Early diagnosis of wound colonisation or 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 aeruginosaresults in high morbidity and mortality rates, is inherently resistant to common antibiotics, and is increasingly being isolated as a nosocomial pathogen. We developed three PCR-based methods to detect and quantifyP aeruginosain wound biopsy samples: conventional PCR, enzyme-linked immunosorbent assay (ELISA)-PCR, and RTD-PCR with rapid thermal cycling (LightCycler™technology), all based on the amplification of the outer membrane lipoprotein geneoprL. We compared the efficacy of these methods to bacterial culture by quantitatively measuring levels ofP aeruginosain serial dilutions, in reconstituted skin samples and 21 burn wound biopsy samples.