TY - JOUR
T1 - A multicenter study of viable PCR using propidium monoazide to detect Legionella in water samples
AU - Scaturro, Maria
AU - Fontana, Stefano
AU - Dell'eva, Italo
AU - Helfer, Fabrizia
AU - Marchio, Michele
AU - Stefanetti, Maria Vittoria
AU - Cavallaro, Mario
AU - Miglietta, Marilena
AU - Montagna, Maria Teresa
AU - De Giglio, Osvalda
AU - Cuna, Teresa
AU - Chetti, Leonarda
AU - Sabattini, Maria Antonietta Bucci
AU - Carlotti, Michela
AU - Viggiani, Mariagabriella
AU - Stenico, Alberta
AU - Romanin, Elisa
AU - Bonanni, Emma
AU - Ottaviano, Claudio
AU - Franzin, Laura
AU - Avanzini, Claudio
AU - Demarie, Valerio
AU - Corbella, Marta
AU - Cambieri, Patrizia
AU - Marone, Piero
AU - Rota, Maria Cristina
AU - Bella, Antonino
AU - Ricci, Maria Luisa
PY - 2016/1/14
Y1 - 2016/1/14
N2 - 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.
AB - 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.
KW - Culture
KW - Legionella
KW - Propidium monoazide
KW - Quantitative PCR
KW - Viable PCR
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U2 - 10.1016/j.diagmicrobio.2016.04.009
DO - 10.1016/j.diagmicrobio.2016.04.009
M3 - Article
AN - SCOPUS:84964286029
JO - Diagnostic Microbiology and Infectious Disease
JF - Diagnostic Microbiology and Infectious Disease
SN - 0732-8893
ER -