Open Access Research article

Detection of Legionella by quantitative-polymerase chain reaction (qPCR) for monitoring and risk assessment

Louise H Krøjgaard12*, Karen A Krogfelt1, Hans-Jørgen Albrechtsen2 and Søren A Uldum1*

  • * Corresponding authors: Louise H Krøjgaard lkd@ssi.dk - Søren A Uldum su@ssi.dk

  • † Equal contributors

Author Affiliations

1 Dept. of Microbiological Surveillance and Research, Statens Serum Institut, Ørestads Boulevard 5, 2300 Copenhagen S, Denmark

2 Dept. of Environmental Engineering, Technical University of Denmark, Miljoevej bygning 113, 2800 Kgs. Lyngby, Denmark

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BMC Microbiology 2011, 11:254  doi:10.1186/1471-2180-11-254

Published: 21 November 2011

Abstract

Background

Culture and quantitative polymerase chain reaction (qPCR) assays for the detection of Legionella were compared on samples from a residential area before and after two interventions. A total of 84 samples were collected from shower hoses and taps as first flush samples and at constant temperature. Samples were grouped according to the origin of the sample, a) circulation water b) water from empty apartments c) water from shower hoses. The aims were to investigate the usefulness of qPCR compared to culture for monitoring remedial actions for elimination of Legionella bacteria and as a tool for risk assessment.

Results

In water collected from the apartments Legionella spp were detected by qPCR in the concentration range from LOQ to 9.6*105GU/L while L. pneumophila were detected in a range from LOQ to 6.8*105 GU/L. By culturing, the legionellae were detected in the range from below detection limit (> 10 CFU/L) to 1.6*106 CFU/L. In circulating water and in first flush water from shower hoses, culture and qPCR showed the same tendencies. The overall correlation between the bacteria number detected by culture and the two developed qPCR assays (L. spp and L. pneumophila) was relatively poor (r2 = 0.31 for culture and Legionella spp. assay, r2 = 0.20 for culture and L. pneumophila assay).

Conclusion

Detection by qPCR was suitable for monitoring changes in the concentration of Legionella but the precise determination of bacteria is difficult. Risk assessment by qPCR only on samples without any background information regarding treatment, timing, etc is dubious. However, the rapid detection by qPCR of high concentrations of Legionella - especially Legionella pneumophila - is valuable as an indicator of risk, although it may be false positive compared to culture results. On the other hand, the detection of a low number of bacteria by qPCR is a strong indication for the absence of risk.