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DNA Mimics for the Rapid Identification of Microorganisms by Fluorescence in situ Hybridization (FISH)
IBB - Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar 4710-057, Braga, Portugal
Environmental Healthcare Unit, School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton SO16 7PX, UK
* Author to whom correspondence should be addressed.
Received: 2 April 2008; in revised form: 24 September 2008 / Accepted: 6 October 2008 / Published: 20 October 2008
Abstract: Fluorescence in situ hybridization (FISH) is a well-established technique that is used for a variety of purposes, ranging from pathogen detection in clinical diagnostics to the determination of chromosomal stability in stem cell research. The key step of FISH involves the detection of a nucleic acid region and as such, DNA molecules have typically been used to probe for the sequences of interest. However, since the turn of the century, an increasing number of laboratories have started to move on to the more robust DNA mimics methods, most notably peptide and locked nucleic acids (PNA and LNA). In this review, we will cover the state-of-the-art of the different DNA mimics in regard to their application as efficient markers for the presence of individual microbial cells, and consider their potential advantages and pitfalls. Available PNA probes are then reassessed in terms of sensitivity and specificity using rRNA databases. In addition, we also attempt to predict the applicability of DNA mimics in well-known techniques attempting to detect in situ low number of copies of specific nucleic acid sequences such as catalyzed reporter deposition (CARD) and recognition of individual genes (RING) FISH.
Keywords: FISH; DNA mimics; PNA; LNA; molecular diagnostics
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Cite This Article
MDPI and ACS Style
Cerqueira, L.; Azevedo, N.F.; Almeida, C.; Jardim, T.; Keevil, C.W.; Vieira, M.J. DNA Mimics for the Rapid Identification of Microorganisms by Fluorescence in situ Hybridization (FISH). Int. J. Mol. Sci. 2008, 9, 1944-1960.
Cerqueira L, Azevedo NF, Almeida C, Jardim T, Keevil CW, Vieira MJ. DNA Mimics for the Rapid Identification of Microorganisms by Fluorescence in situ Hybridization (FISH). International Journal of Molecular Sciences. 2008; 9(10):1944-1960.
Cerqueira, Laura; Azevedo, Nuno F.; Almeida, Carina; Jardim, Tatiana; Keevil, Charles W.; Vieira, Maria J. 2008. "DNA Mimics for the Rapid Identification of Microorganisms by Fluorescence in situ Hybridization (FISH)." Int. J. Mol. Sci. 9, no. 10: 1944-1960.