Abstract:
AIMS:A method for rapid and simultaneous detection, identification and enumeration of specific micro-organisms using Peptide Nucleic Acid (PNA) probes is presented. METHODS AND RESULTS:The method is based on a membrane filtration technique. The membrane filter was incubated for a short period of time. The microcolonies were analysed by in situ hybridization, using peroxidase-labelled PNA probes targeting a species-specific rRNA sequence, and visualized by a chemiluminescent reaction. Microcolonies were observed as small spots of light on film, thereby providing simultaneous detection, identification and enumeration. The method showed 95-100% correlation to standard plate counts along with definitive identification due to the specificity of the probe. CONCLUSION:Using the same protocol, results were generated approximately three times faster than culture methods for Gram-positive and -negative bacterial species and yeast species. SIGNIFICANCE AND IMPACT OF THE STUDY:The method is an improvement on the current membrane filtration technique, providing rapid determination of the level of specific pathogens, spoilage or indicator micro-organisms.
journal_name
J Appl Microbioljournal_title
Journal of applied microbiologyauthors
Perry-O'Keefe H,Stender H,Broomer A,Oliveira K,Coull J,Hyldig-Nielsen JJdoi
10.1046/j.1365-2672.2001.01230.xsubject
Has Abstractpub_date
2001-02-01 00:00:00pages
180-9issue
2eissn
1364-5072issn
1365-2672pii
jam1230journal_volume
90pub_type
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