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Chapter 105 : Suitability of Peptide Nucleic Acid Probes for Detection of in Mains Drinking Water Supplies

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Suitability of Peptide Nucleic Acid Probes for Detection of in Mains Drinking Water Supplies, Page 1 of 2

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Abstract:

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 PNAs are synthetic molecules in which the sugar phosphate backbone has been replaced by 2-aminoethyl-glycine.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 and only , 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 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.

Citation: A. Wilks S, Keevil C. 2006. Suitability of Peptide Nucleic Acid Probes for Detection of in Mains Drinking Water Supplies, p 442-445. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch105

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Figures

Image of FIGURE 1
FIGURE 1

A coupon cut from a cast iron pipe from a mains supply drinking water system. The coupon is approximately 4 cm × 2 cm and has considerable corrosion deposits (up to 2 cm in height), illustrating the problems associated with such samples. The pipe was laid in 1940 in an urban area and had been scraped and bitumen lined in 1979. Pipe supplied by a leading European water company.

Citation: A. Wilks S, Keevil C. 2006. Suitability of Peptide Nucleic Acid Probes for Detection of in Mains Drinking Water Supplies, p 442-445. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch105
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Image of FIGURE 2
FIGURE 2

Coupon cut from a mains supply drinking water system, followed by hybridization with the -specific PNA probe (PLPNE620). (A) EDIC image showing the surface of the pipe. (B) Tetramethyl-rhodamine isocyanate channel showing labeled bacteria on pipe surface. Magnification × 1000.

Citation: A. Wilks S, Keevil C. 2006. Suitability of Peptide Nucleic Acid Probes for Detection of in Mains Drinking Water Supplies, p 442-445. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch105
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References

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