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Author(s): Susana P. Lopes
Daniel T. Carvalho
Maria O. Pereira
Nuno F. Azevedo
Title: Discriminating Typical and Atypical Cystic Fibrosis-Related Bacteria by Multiplex PNA-FISH
Issue Date: 2017
Abstract: This study aims to report the development of peptidenucleic acid (PNA) probes to specifically detect the cystic fibrosis(CF)-associated traditional and atypical species Pseudomonasaeruginosa and Inquilinus limosus, respectively. PNA probes weredesigned in silico, developed and tested in smears prepared inphosphate-buffer saline (PBS), and in artificial sputum medium(ASM). A multiplex fluorescent in situ hybridization (FISH)approach using the designed probes was further validated inartificially contaminated clinical sputum samples and also appliedin polymicrobial 24 h-old biofilms involving P. aeruginosa,I. limosus, and other CF-related bacteria. Both probes showedhigh predictive and experimental specificities and sensitivities. Themultiplex PNA-FISH assay, associated with non-specific staining,was successfully adapted in the clinical samples and in biofilms ofCF-related bacteria, allowing differentiating the communitymembers and inferring about microbial-microbial interactionswithin the consortia. This study revealed the great potential of PNAFISH as a diagnostic tool to discriminate between classical and lesscommon CF-associated bacteria, being suitable to further describespecies-dependent prevention strategies and deliver more effectivetarget control therapeutics.
Related Information: info:eu-repo/grantAgreement/Autoriadade de Gestão do Programa Operacional Regional do Norte/Programas Integrados de IC&DT/NORTE-07-0124-FEDER-000025/(Bio) Chemical Engineering: Multi-Scale Approaches for Sustainable Environment and Health/LEPAE/CEFT - RL2
info:eu-repo/grantAgreement/FCT - Fundação para a Ciência e Tecnologia/Projectos de Investigação Clínica/PIC/IC/82815/2007/Desenvolvimento e aplicação de mímicos de DNA para a rápida identificação de microrganismos patogénicos/DNA mimics
info:eu-repo/grantAgreement/FCT - Fundação para a Ciência e Tecnologia/Projetos Estratégicos/UID/EQU/00511/2013 - POCI-01-0145-FEDER-006939/Laboratório de Engenharia de Processos, Ambiente, Biotecnologia e Energia/LEPABE
Document Type: Artigo em Revista Científica Internacional
Rights: restrictedAccess
Appears in Collections:FEUP - Artigo em Revista Científica Internacional

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