Rapid pulsed-field gel electrophoresis protocol for Legionella pneumophila typing

Massimiliano Orsini, Rosarita Amore, Bianca Maria Pietrangeli, Daniela Anastasi, Walter Ricciardi, Stefania Boccia

Research output: Contribution to journalArticle

Abstract

Background: Genomic DNA patterns generated by pulsed-field gel electrophoresis (PGFE) are highly specific for different strains of an organism and have significant value in epidemiologic investigations of infectious disease outbreaks. A disadvantage of PFGE is that the procedure requires up to 6 days to complete. Methods: We developed a rapid PFGE protocol for subtyping Legionella pneumophila isolates based on the standardized protocol currently used. Various combinations of reaction conditions (e.g., lysis time and temperature, restriction enzyme concentration) and electrophoresis parameters were applied to devise a simple and rapid PFGE protocol that could also be used for frozen bacteria. Results: PFGE analysis of Legionella pneumophila isolates can be completed in 26 hours using this protocol compared to 6 days for the conventional one. Conclusions: We successfully applied a rapid PFGE protocol for Legionella pneumophila typing and comparison of the patterns obtained from the rapid compared with the conventional method showed that the rapid protocol gave identical and highly reproducible results.

Original languageEnglish
Pages (from-to)143-148
Number of pages6
JournalItalian Journal of Public Health
Volume5
Issue number2
Publication statusPublished - 2008

Keywords

  • Legionella pneumophila
  • Molecular typing
  • PFGE

ASJC Scopus subject areas

  • Public Health, Environmental and Occupational Health
  • Health Policy
  • Epidemiology
  • Community and Home Care

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    Orsini, M., Amore, R., Pietrangeli, B. M., Anastasi, D., Ricciardi, W., & Boccia, S. (2008). Rapid pulsed-field gel electrophoresis protocol for Legionella pneumophila typing. Italian Journal of Public Health, 5(2), 143-148.