Author(s):
Lopes, Susana P. ; Carvalho, Daniel T. ; Pereira, Maria Olívia ; Azevedo, Nuno F.
Date: 2017
Persistent ID: http://hdl.handle.net/1822/44751
Origin: RepositóriUM - Universidade do Minho
Project/scholarship:
info:eu-repo/grantAgreement/FCT/5876/147337/PT;
info:eu-repo/grantAgreement/FCT/5876/147284/PT;
info:eu-repo/grantAgreement/FCT/5876-PPCDTI/126270/PT
;
info:eu-repo/grantAgreement/FCT/COMPETE/126270/PT;
Subject(s): cystic fibrosis microbiome; Fluorescence in situ hybridization; Inquilinus limosus; PNA probe; Pseudomonas aeruginosa; Science & Technology
Description
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 and in artificial sputum medium. A multiplex fluorescent in situ hybridization (FISH) approach using the probes was further validated in artificially contaminated clinical sputum samples and also applied to polymicrobial 24h-old biofilms involving the aforementioned and other CF-related bacteria. Both probes showed high predictive and experimental specificities/sensitivities. The multiplex PNA-FISH, 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 PNA-FISH as a diagnostic tool to discriminate between classical and emerging CF-associated bacteria, being suitable to further describe species-dependent prevention strategies and deliver more effective target control therapeutics.
Contract grant sponsor: FCT, COMPETE 2020, FEDER, PT2020Contract grant number: UID/BIO/04469/2013Contract grant sponsor: FCT/MEC, FEDER, PT2020Contract grant number: UID/EQU/00511/2013-LEPABEContract grant sponsor: COMPETE 2020Contract grant number: POCI-01-0145-FEDER-006684Contract grant sponsor: FCTContract grant number: RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462)Contract grant sponsor: QREN, FEDER, ON2Contract grant number: NORTE-07-0124-FEDER-000027Contract grant sponsor: FEDER, ON2, FCT/MECContract grant number: NORTE-07-0124-FEDER-000025Contract grant sponsor: COST-Action TD1004
