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RESEARCH PAPER
Efficacy of the detection of Legionella in hot and cold water samples by culture and PCR. I. Standardization of methods
1
Department of Zoonoses, Institute of Rural Health, Lublin, Poland
2
Department of Water and Soil Safety, Institute of Rural Health, Lublin, Poland
Corresponding author
Ann Agric Environ Med. 2012;19(2):289-293
KEYWORDS
ABSTRACT
The aim of the present study was: – to compare methods for concentration and isolation of Legionella DNA from water; – to examine the efficacy of various modifications of PCR test (PCR, semi-nested PCR, and real-time PCR) for the detection of known numbers of Legionella pneumophila in water samples artificially contaminated with the strain of this bacterium and in randomly selected samples of environmental water, in parallel with examination by culture. It was found that filtration is much more effective than centrifugation for the concentration of DNA in water samples, and that the Qiamp DNA Mini-Kit is the most efficient for isolation of Legionella DNA from water. The semi-nested PCR and real-time PCR proved to be the most sensitive methods for detection of Legionella DNA in water samples. Both PCR modifications showed a high correlation with recovery of Legionella by culture (p<0.01), while no correlation occurred between the results of one-stage PCR and culture (p>0.1).
REFERENCES (17)
1.
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.
2.
Dussere E, Ginevra C, Hallier-Soulier S, Vandenesch F, Festoc G, Etienne J, Jarraud S, Molmeret M. A PCR-based method for monitoring Legionella pneumophila in water samples detects viable but noncultivable legionellae that can recover their cultivability. Appl Environ Microbiol. 2008; 74: 4817-4824.
3.
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.
4.
Szénási Z, Endo T, Yagita K, Veréb I, Nagy E. Epidemiology and laboratory diagnostics of legionellae. Orv Hetil. 2001; 142: 1035-1043.
5.
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.
6.
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.
7.
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.
8.
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.
9.
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.
10.
Ahmadinejad M, Shakibaie MR, Shams K, Khalili M. Detection of Legionella pneumophila in cooling water systems of hospitals and nursing homes of Kerman City, Iran by semi-nested PCR. Int J Biol Life Sci. 2011; 7: 2.
11.
Delgado-Viscogliosi P, Solignac L, Delattre JM. Viability PCR, a culture-independent method for rapid and selective quantification of viable Legionella pneumophila cells in environmental water samples. Appl Environ Microbiol. 2009; 75: 3502-3512.
12.
Fittipaldi M, Codony F, Adrados B, Camper AK, Morató J. Viable real-time PCR in environmental samples: can all data be interpreted directly? Microb Ecol. 2011; 61: 7-12.
13.
Atlas RM, Parks LC. Handbook of Microbiological Media. CRC Press, Boca Raton 1993.
14.
Maleszewska J, Krogulska B, Bielecka Z, Stypułkowska-Misiurewicz H. Presence of Yersinia organism in the water from private wells. Roczn PZH 1988; 39: 396-403.
15.
Matuszewska R, Krogulska B. Detection and isolation of bacteria of genus Legionella from water environment. Roczn PZH 2000; 51: 183-190.
16.
Stypułkowska-Misiurewicz H, Krogulska B, Pancer K, Matuszewska R. Legionella sp. – investigation of human infection and detection in environmental water. Roczn PZH 2001; 52: 1-18 (in Polish).
17.
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.
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