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Applied and Environmental Microbiology, January 2006, p. 848-853, Vol. 72, No. 1
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.1.848-853.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Fluorescence In Situ Hybridization Using Peptide Nucleic Acid Probes for Rapid Detection of Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis in Potable-Water Biofilms

Markku J. Lehtola,1,2* Eila Torvinen,2 Ilkka T. Miettinen,2 and C. William Keevil1

Environmental Healthcare Unit, School of Biological Sciences, University of Southampton, Southampton SO16 7PX, United Kingdom,1 Laboratory of Environmental Microbiology, National Public Health Institute, P.O. Box 95, FI-70701 Kuopio, Finland2

Received 30 May 2005/ Accepted 28 October 2005

Here, we present for the first time a high-affinity peptide nucleic acid (PNA) oligonucleotide sequence for detecting Mycobacterium avium bacteria, including the opportunistically pathogenic subspecies M. avium subsp. avium, M. avium subsp. paratuberculosis, and M. avium subsp. silvaticum, by the fluorescence in situ hybridization (FISH) method. There is evidence that M. avium subsp. avium especially is able to survive and grow in drinking-water biofilms and possibly transmit via drinking water. The designed PNA probe (MAV148) specificity was tested with several bacterial species, including other mycobacteria and mycolic acid-containing bacteria. From the range of bacterial strains tested, only M. avium subsp. avium and M. avium subsp. paratuberculosis strains were hybridized. The PNA FISH method was applied successfully to detect M. avium subsp. avium spiked in water samples and biofilm established within a Propella biofilm reactor fed with potable water from a distribution supply.

* Corresponding author. Mailing address: National Public Health Institute, P.O. Box 95, FI-70701 Kuopio, Finland. Phone: 358 17 201371. Fax: 358 17 201155. E-mail: Markku.Lehtola{at}ktl.fi

Applied and Environmental Microbiology, January 2006, p. 848-853, Vol. 72, No. 1
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.1.848-853.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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