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

• Type: Article in International Scientific Journal
• Date: 2014

Authors

Carina Almeida (Author) Other; Diogo Constante (Author) Other; André Ferreira (Author) Other; Cerqueira, L. (Author) FEUP; Maria J. Vieira (Author) Other; Nuno F. Azevedo (Author) FEUP

Journal

Title: Future Microbiology

Vol. 9 No. 10

Pages: 1131-1142

ISSN: 1746-0913

Publisher: Future Medicine Ltd.

Indexing

ISI Web of Knowledge - 0 Citations

ISI Web of Science

Scopus - 0 Citations

Scientific classification

FOS: Natural sciences > Biological sciences

Associated Projects

UI0511 - PROJECTO ESTRATÉGICO - UI 511 - 2013-2014

Other information

Authenticus ID: 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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 12