Comparative Analysis of Infrequent-Restriction-Site PCR and Pulsed-Field Gel Electrophoresis for Epidemiological Typing of Legionella pneumophila Serogroup 1 Strains

Author:

Riffard Serge1,Presti François Lo1,Vandenesch François1,Forey Françoise1,Reyrolle Monique1,Etienne Jerome1

Affiliation:

1. Centre National de Référence des Legionella, Laboratoire de Bactériologie, UPRES EA1655, Faculté de Médecine R.T.H. Laënnec, 69372 Lyon Cedex 08, France

Abstract

ABSTRACT Two methods were compared for the analysis of 48 unrelated and epidemiologically related Legionella pneumophila serogroup 1 isolates. These are the infrequent-restriction-site PCR (IRS-PCR) assay with adapters designed for Xba I and Pst I restriction sites and the pulsed-field gel electrophoresis (PFGE) analysis determined after DNA restriction with Sfi I. Both methods demonstrated a high level of discrimination with a similar capacity for differentiating 23 of the 24 unrelated isolates. PFGE analysis and IRS-PCR assay were both able to identify epidemiologically related isolates of L. pneumophila from three outbreaks. Hence, IRS-PCR assay appears to be a reproducible (intergel reproducibility, 100%) and discriminative (discriminatory index, ≥0.996) tool for typing of Legionella . Compared to PFGE, however, IRS-PCR presented an advantage through ease of performance and with attributes of rapidity and sensitivity of target DNA.

Publisher

American Society for Microbiology

Subject

Microbiology (medical)

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