Premium
Filter‐based PNA in situ hybridization for rapid detection, identification and enumeration of specific micro‐organisms
Author(s) -
PerryO’Keefe H.,
Stender H.,
Broomer A.,
Oliveira K.,
Coull J.,
HyldigNielsen J.J.
Publication year - 2001
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.2001.01230.x
Subject(s) - library science , identification (biology) , biology , computer science , ecology
H. PERRY‐O’KEEFE, H. STENDER, A. BROOMER, K. OLIVEIRA, J. COULL AND J.J. HYLDIG‐NIELSEN. 2001 . 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. Conclusions: 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.