A new colorimetric peptide nucleic acid-based assay for the specific detection of bacteria

• Tipo: Artigo em Revista Científica Internacional
• Data: 2014

Autores

Carina Almeida (Autor) Outra; Diogo Constante (Autor) Outra; André Ferreira (Autor) Outra; Laura Cerqueira (Autor) Outra; Maria J. Vieira (Autor) Outra; Nuno F. Azevedo (Autor) FEUP

Revista

Título: Future Microbiology

Vol. 9 Nº 10

Páginas: 1131-1142

ISSN: 1746-0913

Editora: Future Medicine Ltd.

Indexação

ISI Web of Knowledge - 0 Citações

ISI Web of Science

Scopus - 0 Citações

Classificação Científica

FOS: Ciências exactas e naturais > Ciências biológicas

Projetos Associados

UI0511 - PROJECTO ESTRATÉGICO - UI 511 - 2013-2014

Outras Informações

ID Authenticus: P-009-YX9

DOI: 10.2217/fmb.14.68

Abstract (EN): 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.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 12