Efficacy of the detection of Legionella in hot and cold water samples by culture and PCR. II. Examination of native samples from various sources
More details
Hide details
Department of Water and Soil Safety, Institute of Rural Health, Lublin, Poland
Department of Zoonoses, Institute of Rural Health, Lublin, Poland
Corresponding author
Nimfa Maria Stojek   

Department of Water and Soil Safety, Institute of Rural Health, Lublin, Poland
Ann Agric Environ Med. 2012;19(2):295-298
A total of 123 water samples were examined in parallel by culture and semi-nested PCR for the presence of Legionella. They comprised: 35 samples of hot water distributed by the urban municipal water supply system (MWSS) taken in institutions, 45 samples of hot water distributed by urban MWSS taken in dwellings, 27 samples of cold water distributed by rural MWSS taken in dwellings, and 16 samples of cold well water taken in rural areas. The greatest frequency of the isolation of Legionella by culture (88.6%) was recorded in the samples of hot water from the urban institutions, having been greater compared to all other sources (p<0.001). The frequency of Legionella isolation from hot water in urban dwellings (28.9%) was significantly greater compared to the combined value (2.3%) for cold water from rural MWSS and wells (p<0.001). Strains belonging to Legionella pneumophila serogroups 2-14 predominated in the examined samples, while strains of L. pneumophila serogroup 1 and strains of Legionella spp. (other than L. pneumophila) were 3-fold less numerous. The rates of positive findings in the semi-nested PCR (stage 2) were greater than culture isolations in all kinds of samples, except for urban institutions. The correlation between the culture and PCR results was positive for samples of hot water from urban MWSS (p<0.01), but not for samples of cold water from rural MWSS and wells (p>0.5). A significant correlation was found between rates of PCR-positive results and numbers of Legionella pneumophila serogroups 2-14 strains, but not for other Legionella serogroups or species. In conclusion, our results support the opinion that though PCR cannot be a substitute for the isolation of Legionella by culture, it could be regarded as an useful complementary method.
Weissenberger CA, Cazalet C, Buchrieser C. Legionella pneumophila – a human pathogen that co-evolved with fresh water protozoa. Cell Molec Life Sci. 2007; 64: 432-448.
Stojek NM, Dutkiewicz J. Legionella and other Gram-negative bacteria in potable water from various rural and urban sources. Ann Agric Environ Med. 2006; 13: 323-335.
Rozporządzenie Ministra Zdrowia z dnia 13 marca 2007 r. w sprawie jakości wody przeznaczonej do spożycia przez ludzi. Dz. U. Nr 61, poz. 417, Warsaw 2007.
Wellinghausen N, Frost C, Marre R. Detection of legionellae in hospital water samples by quantitative real-time LightCycler PCR. Appl Environ Microbiol. 2001; 67: 3985-3993.
Szénási Z, Endo T, Yagita K, Veréb I, Nagy E. Epidemiology and laboratory diagnostics of legionellae. Orv Hetil. 2001; 142: 1035-1043.
Joly P, Falconnet P-A, André J, Weill N, Reyrolle M, Vandenesch F, Maurin M, Etienne J, Jarraud S. Quantitative real-time Legionella PCR for environmental water samples: data interpretation. Appl Environ Microbiol. 2006; 72: 2801-2808.
Yaradou DF, Hallier-Soulier S, Moreau S, Poty F, Hillion Y, Reyrolle M, André J, Festoc G, Delabre K, Vandenesch F, Etienne J, Jarraud S. Integrated real-time PCR for detection and monitoring of Legionella pneumophila in water systems. Appl Environ Microbiol. 2007; 73: 1452-1456.
Morio F, Corvec S, Caroff N, Le Gallou F, Drugeon H, Reynaud A. Real-time PCR assay for the detection and quantification of Legionella pneumophila in environmental water samples: utility for daily practice. Int J Hyg Environ Health. 2008; 211: 403-411.
Guillemet TA, Lévesque B, Gauvin D, Brousseau N, Giroux JP, Cantin P. Assessment of real-time PCR for quantification of Legionella spp. in spa water. Lett Appl Microbiol. 2010; 51: 639-644.
Lee JV, Lai S, Exner M, Lenz J, Gaia V, Casati S, Hartemann P, Lück C, Pangon B, Ricci ML, Scaturro M, Fontana S, Sabria M, Sánchez I, Assaf S, Surman-Lee S. An international trial of quantitative PCR for monitoring Legionella in artificial water systems. Appl Microbiol. 2011. doi: 10.1111/j.1365-2672.2011.04957.x.
Wójcik-Fatla A, Stojek NM, Dutkiewicz J. Efficacy of the detection of Legionella in hot and cold water samples by culture and PCR: I. Standardization methods. Ann Agric Environ Med. 2012; 19(2): 289-293.
Atlas RM, Parks LC. Handbook of Microbiological Media. CRC Press, Boca Raton 1993.
Yanez MA, Barberá VM, Catalán V. Validation of a new seminested PCR-based detection method for Legionella pneumophila. J Microbiol Methods. 2007; 70: 214-217.
Journals System - logo
Scroll to top