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TitleA new colorimetric peptide nucleic acid-based assay for the specific detection of bacteria
Author(s)Almeida, Carina
Constante, Diogo
Ferreira, André
Cerqueira, L.
Vieira, M. J.
Azevedo, N. F.
KeywordsColorimetric detection
Issue date2014
PublisherFuture Medicine
JournalFuture microbiology
CitationAlmeida, Carina; Constante, Diogo; Ferreira, André; Cerqueira, L.; Vieira, M. J.; Azevedo, N. F., A new colorimetric peptide nucleic acid-based assay for the specific detection of bacteria. Future Microbiology, 9(10), 1131-1142, 2014
Abstract(s)Aim: Developments on synthetic molecules, such as peptide nucleic acid (PNA), make FISH procedures more robust for microbial identification. Fluorochromes use might hinder a broader implementation of PNA-FISH, but colorimetric applications are inexistent so far. Methods: A biotin-labeled eubacteria probe was used to develop a colorimetric PNA-in situ hybridization (ISH) assay. An enzymatic-conjugate, targeting biotin, was introduced. The procedure was optimized and evaluated regarding sensitivity, specificity and detection limit. Results: Results have shown strong ISH signals. The method was specific, but permeabilization problems were observed for Gram-positive bacteria. Detection limit was 5 × 107 CFU/ml, limiting current applications to pre-enriched samples. Conclusion: The PNA-ISH procedure described here is a simple alternative to other detection methods, and is also the base for the development of other PNA colorimetric systems.
AccessRestricted access (UMinho)
Appears in Collections:CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series

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