TY - JOUR AB - Legionella pneumophila (L. pneumophila) is a harmful pathogen often found in water systems. In hospitals, the absence of L. pneumophila in water systems is mandatory by law, therefore, frequent and effective monitoring of water is of fundamental importance. Molecular methods based on reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) have been proposed for the detection of L. pneumophila, however, the sensitivity and accuracy of these methods have not been validated yet. Therefore, it is important to evaluate other strategies able to overcome the limits of culture‑based and RT‑qPCR methods. On these bases, we compared the sensitivity and accuracy of droplet digital PCR (ddPCR) and RT‑qPCR in water samples with known concentrations of L. pneumophila and in an in vitro model of water heat treatments. ddPCR showed a higher sensitivity rate and accuracy compared to RT‑qPCR in detecting low bacterial load. In addition, ddPCR is not affected by the presence of fragmented DNA and showed higher accuracy than RT‑qPCR in monitoring the efficacy of heat shock treatments. In conclusion, ddPCR represents an innovative strategy to effectively detect L. pneumophila in water samples. Thanks to its high robustness, ddPCR could be applied also for the detection of L. pneumophila in patients with suspected legionellosis. AD - Epidemiology Unit, IRCCS Istituto Nazionale Tumori ‘Fondazione G. Pascale’, I‑80131 Naples, Italy Department of Biomedical and Biotechnological Sciences, Section of General Pathology, University of Catania, I‑95123 Catania, Italy Department of Biomedical and Biotechnological Sciences, Section of Microbiology, University of Catania, I‑95123 Catania, Italy Department of Biomedical and Biotechnological Sciences, Section of Biochemistry, University of Catania, I‑95123 Catania, Italy Department of Medical Sciences, Surgical and Advanced Technologies, GF Ingrassia, University of Catania, I‑95123 Catania, Italy Laboratory of Clinical Virology, Medical School, University of Crete, 71003 Heraklion, Greece AU - Falzone,Luca AU - Gattuso,Giuseppe AU - Lombardo,Cinzia AU - Lupo,Gabriella AU - Grillo,Caterina,Maria AU - Spandidos,Demetrios,A. AU - Libra,Massimo AU - Salmeri,Mario DA - 2020/11/01 DO - 10.3892/ijmm.2020.4724 EP - 1782 IS - 5 JO - Int J Mol Med KW - Legionella pneumophila ddPCR RT‑qPCR diagnosis sensitivity molecular biology PY - 2020 SN - 1107-3756 1791-244X SP - 1777 ST - Droplet digital PCR for the detection and monitoring of Legionella pneumophila T2 - International Journal of Molecular Medicine TI - Droplet digital PCR for the detection and monitoring of Legionella pneumophila UR - https://doi.org/10.3892/ijmm.2020.4724 VL - 46 ER -