Article

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

Environmental Healthcare Unit, School of Biological Sciences, University of Southampton, Southampton SO16 7PX, United Kingdom.
Applied and Environmental Microbiology (Impact Factor: 3.95). 02/2006; 72(1):848-53. DOI: 10.1128/AEM.72.1.848-853.2006
Source: PubMed

ABSTRACT 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.

Download full-text

Full-text

Available from: Charles William Keevil, Jul 01, 2015
0 Followers
 · 
85 Views
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Scientific imaging represents an important and accepted research tool for the analysis and understanding of complex natural systems. Apart from traditional microscopic techniques such as light and electron microscopy, new advanced techniques have been established including laser scanning microscopy (LSM), magnetic resonance imaging (MRI) and scanning transmission X-ray microscopy (STXM). These new techniques allow in situ analysis of the structure, composition, processes and dynamics of microbial communities. The three techniques open up quantitative analytical imaging possibilities that were, until a few years ago, impossible. The microscopic techniques represent powerful tools for examination of mixed environmental microbial communities usually encountered in the form of aggregates and films. As a consequence, LSM, MRI and STXM are being used in order to study complex microbial biofilm systems. This mini review provides a short outline of the more recent applications with the intention to stimulate new research and imaging approaches in microbiology.
    FEMS Microbiology Ecology 04/2010; 72(1):1-21. DOI:10.1111/j.1574-6941.2010.00837.x · 3.88 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: A fluorescently labelled peptide nucleic acid (PNA) probe has been applied for the in situ detection of Helicobacter pylori in drinking water biofilms. The method was originally applied to real pipe samples removed from a drinking water distribution system (DWDS) but the curvature and the heavy fouling of the pipes prevented an accurate detection of the bacterium by epifluorescence microscopy. Therefore, two semi-circular flow cells were placed in a bypass of the DWDS, and coupons with up to 72 days of exposure were regularly sampled and analysed for the presence of H. pylori. In the flat surfaces of the coupons, it was possible to sparsely detect cells exhibiting similar morphology to H. pylori that were emitting the PNA probe fluorescent signal. Coupons were also visualised under the microscope before the hybridisation procedure to serve as negative controls and ensure the validity of the method. This work corroborates the findings already published elsewhere that this bacterium might be present in DWDS biofilms. The method requires, however, highly trained personnel for an accurate detection of the pathogen and will need simplification before being routinely used in standard water analysis laboratories.
    Water Science & Technology 02/2007; 55(8-9):387-93. DOI:10.2166/wst.2007.282 · 1.21 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Helicobacter pylori are Gram-negative micro-aerophilic motile curve rods that inhabit the gastric mucosa of the human stomach. The bacterium chronically infects billions of people worldwide and is one of the most genetically diverse of bacterial species. More than half of the world population in both developed and developing countries are infected with this organism. Infection usually occurs without overt clinical symptoms, particularly in poor communities. If untreated, the infection can last for decades without causing symptoms. In some communities, however, infection with the organism causes peptic and duodenal ulcers, gastritis, duodenitis, and gastric cancers. How H. pylori initially enters the stomach is not known, but contaminated food particles and water are suspected, with the former physically shielding it from stomach acid. Similarly, the route of transmission of this pathogen is unknown. Several reports have suggested the possibility of waterborne transmission as the organism can survive for a few days in fresh cold water, salt water, distilled water, and tap water. Knowledge of the epidemiology and mode of transmission of H. pylori is important to prevent its spread and may be useful in identifying high risk populations.
    Reviews on environmental health 01/2009; 24(1):1-14. DOI:10.1515/REVEH.2009.24.1.1