Please use this identifier to cite or link to this item:
https://hdl.handle.net/10216/104971
Author(s): | Carina Almeida Diogo Constante André Ferreira Laura Cerqueira Maria J. Vieira Nuno F. Azevedo |
Title: | A new colorimetric peptide nucleic acid-based assay for the specific detection of bacteria |
Issue Date: | 2014 |
Abstract: | 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 x 10(7) 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. |
Subject: | Ciências biológicas Biological sciences |
Scientific areas: | Ciências exactas e naturais::Ciências biológicas Natural sciences::Biological sciences |
URI: | https://hdl.handle.net/10216/104971 |
Related Information: | info:eu-repo/grantAgreement/FCT - Fundação para a Ciência e a Tecnologia/Projetos Estratégicos/PEst-C/EQB/UI0511/2013/PROJECTO ESTRATÉGICO - UI 511 - 2013-2014/UI0511 |
Document Type: | Artigo em Revista Científica Internacional |
Rights: | restrictedAccess |
Appears in Collections: | FEUP - Artigo em Revista Científica Internacional |
Files in This Item:
File | Description | Size | Format | |
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104742.1.pdf | Post-Print version | 840.25 kB | Adobe PDF | View/Open |
104742.pdf Restricted Access | Artigo original publicado | 2.69 MB | Adobe PDF | Request a copy from the Author(s) |
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