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Reliability of Pseudomonas aeruginosa semi-automated rep-PCR genotyping in various epidemiological situations

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The purpose of this study was to evaluate the possibility of using a semi-automated repetitive DNA sequences-based polymerase chain reaction (rep-PCR) for typing Pseudomonas aeruginosa isolates. rep-PCR profiles obtained by the DiversiLab® system of 84 P. aeruginosa isolates from distinct epidemiological situations were obtained. rep-PCR groupings were in good agreement with the origin of these isolates. Linked rep-PCR profiles were observed for isolates recovered from a same family of cystic fibrosis (CF) patients, for the etiological agents of clustered cases of nosocomial infections, and for some isolates recovered from a same hospital room. rep-PCR and pulsed-field gel electrophoresis SpeI restricted genomic DNA (PFGE-SpeI) profiles were compared. In a few instances, rep-PCR revealed genetic divergences among isolates of a same group of PFGE-SpeI profiles. These divergences could reflect genetic drifts among closely related isolates, as illustrated by those observed between clinical and environmental isolates of a same group of PFGE-SpeI profiles. The interpretation of such differences will require further studies, but the rep-PCR analysis of P. aeruginosa diversity appeared to be an appropriate method to investigate infra-specific genetic relatedness.

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The authors acknowledge Dr. S. Tigaud at Croix-Rousse Hospital for the initiation of this study and L. Loiseau for her technical assistance.

The authors would like to thank the Cluster “Environnement” of the Rhône-Alpes Region and the CNRS for their financial support of the PAR-MIC technical platform (UMR5557 Microbial Ecology, Villeurbanne, France). This work was partly supported by the Agence Nationale de la Recherche (ANR) SEST 2005 009-01 and CESA 2008 022-01 projects.

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Correspondence to A. Doléans-Jordheim.

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A. Doléans-Jordheim and B. Cournoyer contributed equally to this work.

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Doléans-Jordheim, A., Cournoyer, B., Bergeron, E. et al. Reliability of Pseudomonas aeruginosa semi-automated rep-PCR genotyping in various epidemiological situations. Eur J Clin Microbiol Infect Dis 28, 1105–1111 (2009).

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