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.
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.
Authors: H Stender; A J Broomer; K Oliveira; H Perry-O'Keefe; J J Hyldig-Nielsen; A Sage; J Coull Journal: Appl Environ Microbiol Date: 2001-01 Impact factor: 4.792
Authors: Laura Cerqueira; Ricardo M Fernandes; Rui M Ferreira; Mónica Oleastro; Fátima Carneiro; Catarina Brandão; Pedro Pimentel-Nunes; Mário Dinis-Ribeiro; Céu Figueiredo; Charles W Keevil; Maria J Vieira; Nuno F Azevedo Journal: J Clin Microbiol Date: 2013-04-17 Impact factor: 5.948
Authors: Laura Cerqueira; Nuno F Azevedo; Carina Almeida; Tatiana Jardim; Charles William Keevil; Maria J Vieira Journal: Int J Mol Sci Date: 2008-10-20 Impact factor: 5.923