| Literature DB >> 27133308 |
Maria Scaturro1, Stefano Fontana2, Italo Dell'eva3, Fabrizia Helfer3, Michele Marchio3, Maria Vittoria Stefanetti4, Mario Cavallaro4, Marilena Miglietta4, Maria Teresa Montagna5, Osvalda De Giglio5, Teresa Cuna5, Leonarda Chetti6, Maria Antonietta Bucci Sabattini6, Michela Carlotti6, Mariagabriella Viggiani6, Alberta Stenico7, Elisa Romanin7, Emma Bonanni8, Claudio Ottaviano8, Laura Franzin9, Claudio Avanzini9, Valerio Demarie9, Marta Corbella10, Patrizia Cambieri10, Piero Marone10, Maria Cristina Rota2, Antonino Bella2, Maria Luisa Ricci2.
Abstract
Legionella quantification in environmental samples is overestimated by qPCR. Combination with a viable dye, such as Propidium monoazide (PMA), could make qPCR (named then vPCR) very reliable. In this multicentre study 717 artificial water samples, spiked with fixed concentrations of Legionella and interfering bacterial flora, were analysed by qPCR, vPCR and culture and data were compared by statistical analysis. A heat-treatment at 55 °C for 10 minutes was also performed to obtain viable and not-viable bacteria. When data of vPCR were compared with those of culture and qPCR, statistical analysis showed significant differences (P < 0.001). However, although the heat-treatment caused an abatement of CFU/mL ≤1 to 1 log10 unit, the comparison between untreated and heat-treated samples analysed by vPCR highlighted non-significant differences (P > 0.05). Overall this study provided a good experimental reproducibility of vPCR but also highlighted limits of PMA in the discriminating capability of dead and live bacteria, making vPCR not completely reliable.Entities:
Keywords: Culture; Legionella; Propidium monoazide; Quantitative PCR; Viable PCR
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Year: 2016 PMID： 27133308 DOI： 10.1016/j.diagmicrobio.2016.04.009
Source DB: PubMed Journal: Diagn Microbiol Infect Dis ISSN： 0732-8893 Impact factor: 2.803