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Int. J. Mol. Sci. 2008, 9(10), 1944-1960; doi:10.3390/ijms9101944
Review

DNA Mimics for the Rapid Identification of Microorganisms by Fluorescence in situ Hybridization (FISH)

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Received: 2 April 2008 / Revised: 24 September 2008 / Accepted: 6 October 2008 / Published: 20 October 2008
(This article belongs to the Special Issue Nucleic Acid Derivatives in Emerging Technologies)
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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 FISH; DNA mimics; PNA; LNA; molecular diagnostics
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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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.

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