Assessing the viability of Legionella pneumophila in environmental samples: regarding the filter application of Ethidium Monoazide Bromide

Abstract Purpose Analyses of 34 water samples from 13 healthcare structures revealed how culture method and quantitative PCR (qPCR) often differ in the detection Legionella pneumophila ( Lp ). With these considerations hand, method, Ethidium Monoazide Bromide (EMA) qPCR have all been compared order to detect samples, identify a able speed up procedures, “viable but not cultivable” bacteria (VBNC) exclude non-viable using commercial kit for extraction amplification as well modification protocol. Methods Pure artificially spiked with viable, VBNC ATCC 33152 were analyzed both EMA-qPCR, while ISO 11731-2-2004 was used method. Results Only 35% (12/34) environmental positive methods. regard results showed absence dye toxicity on viable strains an incomplete effectiveness ones. In strains, decrease bacterial DNA recorded function sample dilution EMA concentration. Conclusions Discrepancies between EMA-qPCR observed may be due different causes such membrane-dye interactions, presence interfering compounds sensitivity used. Study significance impact one or more suspected cases nosocomial legionellosis, application rapid molecular only would useful quickly source infection intervene sanitation treatments. However, seeing that our experience pretreatment filter membrane did come expected results, it necessary proceed other experiments and/or dyes. Graphical