Suitability of peptide nucleic acid probes to detect Legionellae in mains drinking water supplies

Wilks, Sandra and Keevil, Charles (2006) Suitability of peptide nucleic acid probes to detect Legionellae in mains drinking water supplies. In, Cianciotto, Nicholas P., Kwaik, Yousef Abu, Edelstein, Paul H., Fields, Barry S., Geary, David F., Harrison, Timothy G., Joseph, Carol A., Ratcliff, Rodney M., Stout, Janet E. and Swanson, Michele S. (eds.) Legionella: State of the Art 30 Years after Its Recognition: Contributions presented at the 6th International Conference on Legionella, Chicago, Illinois, USA, 16-20 October 2005. 6th International Conference on Legionella (16/10/05 - 20/10/05) ASM Press, pp. 442-445. (doi:10.1128/9781555815660.ch105).

Abstract

Legionellae are ubiquitous in the environment. Molecular techniques have been applied, but they too have inherent limitations and rarely provide quantitative data. Consequently, there is a real need for a rapid and direct detection method. Each DNA probe has its own specific requirements, and the hybridization procedure is highly sensitive to changes in temperature, pH, and ionic conditions. This causes problems when working with environmental samples. An alternative is to use peptide nucleic acid (PNA) probes. PNAs are synthetic molecules in which the sugar phosphate backbone has been replaced by 2-aminoethyl-glycine. They exhibit sequence-specific recognition of both DNA and RNA, obeying Watson-Crick hydrogen bonding rules. A study has been undertaken to directly compare the binding efficiency of two published DNA probes against two newly designed PNA probes. The DNA probes tested were LEG226 and LegPNE1, which target all species of Legionella and only Legionella pneumophila, respectively. The thermal stability and chemical resistance of PNA probes make them a valuable tool for use with complex environmental samples and will permit the simultaneous labeling of several target species. It has been shown previously that Legionellae are widespread in potable water systems, but there is no information on population sizes and the proportion associated with biofilm compared to the bulk water. As outbreaks of Legionnaires' disease continue, it is imperative that the authors increase their understanding of the distribution and prevalence of this serious human pathogen.

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Contributors

Author: Sandra Wilks

Author: Charles Keevil

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