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DNA mimics for the rapid identification of microorganisms by fluorescence in situ hybridization (FISH)

DNA mimics for the rapid identification of microorganisms by fluorescence in situ hybridization (FISH)
DNA mimics for the rapid identification of microorganisms by fluorescence in situ hybridization (FISH)
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.
fish, dna mimics, pna, lna, molecular diagnostics
1422-0067
1944-1960
Cerqueira, Laura
61343b41-4742-4057-94e9-2ef4ea145613
Azevedo, Nuno F.
24c4eb52-0c98-443b-881f-7a1449c9ac26
Almeida, Carina
08bcd2c4-bb43-482c-a3ed-517f5d1b9abf
Jardim, Tatiana
31d2ae58-0b9c-4d26-a915-9f52b1a08879
Keevil, Charles William
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Vieira, Maria J.
d972e877-d85b-488c-8b0f-358f79d2fa29
Cerqueira, Laura
61343b41-4742-4057-94e9-2ef4ea145613
Azevedo, Nuno F.
24c4eb52-0c98-443b-881f-7a1449c9ac26
Almeida, Carina
08bcd2c4-bb43-482c-a3ed-517f5d1b9abf
Jardim, Tatiana
31d2ae58-0b9c-4d26-a915-9f52b1a08879
Keevil, Charles William
cb7de0a7-ce33-4cfa-af52-07f99e5650eb
Vieira, Maria J.
d972e877-d85b-488c-8b0f-358f79d2fa29

Cerqueira, Laura, Azevedo, Nuno F., Almeida, Carina, Jardim, Tatiana, Keevil, Charles William and Vieira, Maria J. (2008) DNA mimics for the rapid identification of microorganisms by fluorescence in situ hybridization (FISH). [in special issue: Nucleic Acid Derivatives in Emerging Technologies] International Journal of Molecular Sciences, 9 (10), 1944-1960. (doi:10.3390/ijms9101944). (PMID:19325728)

Record type: Article

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.

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More information

Published date: 20 October 2008
Keywords: fish, dna mimics, pna, lna, molecular diagnostics
Organisations: Centre for Biological Sciences

Identifiers

Local EPrints ID: 209315
URI: https://eprints.soton.ac.uk/id/eprint/209315
ISSN: 1422-0067
PURE UUID: fd809923-95f7-4a11-b1ff-c1a6e123d6d2
ORCID for Charles William Keevil: ORCID iD orcid.org/0000-0003-1917-7706

Catalogue record

Date deposited: 31 Jan 2012 12:09
Last modified: 29 Oct 2019 01:54

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