Confronto tra la tecnica colturale standard e la PCR real-time quantitativa per la identificazione di Legionella pneumophila in campioni di acqua di circuito ospedaliero

Translated title of the contribution: [Autom. eng. transl.] Comparison between standard cultivation technique and quantitative real-time PCR for the identification of Legionella pneumophila in hospital circuit water samples

Stefania Boccia, Patrizia Laurenti, Stefania Bruno, Gianfranco Damiani, Umberto Moscato, Brunella Posteraro, Gianluigi Quaranta, Gualtiero Ricciardi, Emanuele Leoncini, Paolo Di Giannantonio, Sara Vincenti, Federica Boninti, Dario Arzani, Romina Sezzatini, Maria Giovanna Ficarra, Rosarita Amore, Alessia Vecchioni

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

[Autom. eng. transl.] INTRODUCTION: Legionella is a ubiquitous bacterium in aqueous environments. In Europe, approximately 90% of the infections caused by this bacterium are due to L. pneumophila with type 1 serogroup. The standard method used for environmental surveillance is the culture technique, although the method based on the PCR Real-time PCR represents a attractive alternative. The objective of this study is the identification of the threshold value, calculated with Real-time PCR, which reflects the greatest concordance with the values of the conventional microbiological technique. MATERIALS AND METHODS: From 2011 to 2013, 77 water samples were collected in sterile bottles, containing 0.01% sodium thiosulphate to neutralize chlorine residues, and transported at controlled temperature to the laboratories for subsequent analysis. L. pneumophila DNA was extracted and then quantified by Real-time PCR using the iQ-Check Screen kit L. pneumophila (Biorad). RESULTS: Twenty samples (26%) out of 77 analyzed tested positive for the standard culture method. The concentrations of L. pneumophila, determined by PCR as GU / l, were greater than those expressed in CFU / l in 56 samples (73%), minor in only 1 sample (1%), and equal in 20 samples (26 %). A significant correlation between the methods has been reported (ρ = 0.52). CONCLUSIONS: The high sensitivity and negative predictive value detected, make Real-time PCR an ideal screening method to identify and quantify L. pneumophila in environmental water samples. When compared with cultivation methods it has the advantage of providing results in less than 3 hours after water filtration steps and DNA extraction. The Real-time PCR therefore represents an interesting technique complementary to the standard cultivation method.
Translated title of the contribution[Autom. eng. transl.] Comparison between standard cultivation technique and quantitative real-time PCR for the identification of Legionella pneumophila in hospital circuit water samples
Original languageItalian
Title of host publicationATTI del 47° Congresso Nazionale SItI - Comunicazioni brevi
Pages276
Number of pages1
Publication statusPublished - 2014
Event47° Congresso Nazionale SItI - Riccione
Duration: 1 Oct 20144 Oct 2014

Conference

Conference47° Congresso Nazionale SItI
CityRiccione
Period1/10/144/10/14

Keywords

  • PCR real time
  • legionella pneumophila

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