Discriminating Typical and Atypical Cystic Fibrosis-Related Bacteria by Multiplex PNA-FISH

• Type: Article in International Scientific Journal
• Date: 2017

Authors

Susana P. Lopes (Author) Other; Daniel T. Carvalho (Author) Other; Maria O. Pereira (Author) Other; Nuno F. Azevedo (Author) FEUP

Journal

Title: Biotechnology and Bioengineering

Vol. 114 No. 2

Pages: 355-367

ISSN: 0006-3592

Publisher: Wiley-Blackwell

Indexing

ISI Web of Science

Scopus

Associated Projects

LEPAE/CEFT - RL2 - (Bio) Chemical Engineering: Multi-Scale Approaches for Sustainable Environment and Health

DNA mimics - Desenvolvimento e aplicação de mímicos de DNA para a rápida identificação de microrganismos patogénicos

LEPABE - Laboratório de Engenharia de Processos, Ambiente, Biotecnologia e Energia

Other information

DOI: 10.1002/bit.26085

Abstract (EN): This study aims to report the development of peptide nucleic acid (PNA) probes to specifically detect the cystic fibrosis (CF)-associated traditional and atypical species Pseudomonas aeruginosa and Inquilinus limosus, respectively. PNA probes were designed in silico, developed and tested in smears prepared in phosphate-buffer saline (PBS), and in artificial sputum medium (ASM). A multiplex fluorescent in situ hybridization (FISH) approach using the designed probes was further validated in artificially contaminated clinical sputum samples and also applied in polymicrobial 24 h-old biofilms involving P. aeruginosa, I. limosus, and other CF-related bacteria. Both probes showed high predictive and experimental specificities and sensitivities. The multiplex PNA-FISH assay, associated with non-specific staining, was successfully adapted in the clinical samples and in biofilms of CF-related bacteria, allowing differentiating the community members and inferring about microbial-microbial interactions within the consortia. This study revealed the great potential of PNAFISH as a diagnostic tool to discriminate between classical and less common CF-associated bacteria, being suitable to further describe species-dependent prevention strategies and deliver more effective target control therapeutics.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 13