Project/scholarship details


  • Funder

    FCT - Fundação para a Ciência e a Tecnologia, I.P.

  • Funder's country

    Portugal

  • Funding program

    5646-ICCMS

  • Funding amount

    125,677.00 €

  • Start date

    2012-02-01

  • End date

    2015-07-30

Documents


Validation of a fluorescence in situ hybridization method using peptide nucleic...

Cerqueira, L.; Fernandes, R. M.; Ferreira, Rui M.; Oleastro, Mónica; Carneiro, Fátima; Brandão, Catarina; Nunes, Pedro Pimentel; Ribeiro, Mário Dinis

Here, we evaluated a previously established peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) method as a new diagnostic test for Helicobacter pylori clarithromycin resistance detection in paraffin-embedded gastric biopsy specimens. Both a retrospective study and a prospective cohort study were conducted to evaluate the specificity and sensitivity of a PNA-FISH method to determine H. pylori cla...


Rapid detection of urinary tract infections caused by Proteus spp. using PNA-FISH

Almeida, Carina; Azevedo, N. F.; Bento, Joana C.; Cerca, Nuno; Ramos, H.; Vieira, M. J.; Keevil, C. W.

We developed a fluorescence in situ hybridization (FISH) method for the rapid detection of Proteus spp. in urine, using a novel peptide nucleic acid (PNA) probe. Testing on 137 urine samples from patients with urinary tract infections has shown specificity and sensitivity values of 98 % (95 % CI, 93.2–99.7) and 100 % (95 % CI, 80,8–100), respectively, when compared with CHROMagar Orientation medium. Results ind...


Detection of Escherichia coli O157 by Peptide Nucleic Acid Fluorescence In Situ...

Almeida, C.; Sousa, J. M.; Rocha, Rui; Cerqueira, L.; Fanning, S.; Azevedo, N. F.; Vieira, Maria João

Despite the emergence of non-O157 shiga toxin-producing Escherichia coli (STEC) infections, E. coli O157 serotype is still the most commonly identified STEC in world. It causes high morbidity and mortality, and has been responsible for a number of outbreaks in many parts of the world. Various methods have been developed to detect this particular serotype, but standard bacteriological methods remain as the gold ...


Detection and discrimination of microorganisms using Locked Nucleic Acid - Fluo...

Azevedo, A. S.; Pereira, B.; Almeida, Carina; Melo, Luís F.; Wengel, J.; Azevedo, N. F.

The impact of multispecies biofilms on catheter-associated urinary tract infections outcome is still unclear due to the lack of adequate methodologies to discriminate the populations in situ. Employing fluorescence in situ hybridization (FISH) to discriminate the populations in a biofilm, can contribute to the understanding of microorganisms interactions in this structures. Consequently, this information might ...


Optimization of peptide nucleic acid fluorescence in situ hybridization (PNA-FI...

Rocha, Rui; Santos, R.; Madureira, P.; Almeida, Carina; Azevedo, N. F.

Fluorescence in situ hybridization (FISH) appeared in the 1980’s and is nowadays widely used in the field of microbiology. FISH is affected by a wide variety of abiotic and biotic variables and their interplay. This is translated into a wide variability of FISH procedures that can be found in the literature. The aim of this work is to study the effects of pH, probe and dextran sulphate concentration in the FISH...


Optimization of peptide nucleic acid fluorescence in situ hybridization (PNA-FI...

Rocha, Rui; Santos, Rita S.; Pedro, Madureira; Almeida, Carina; Azevedo, Nuno F.

Fluorescence in situ hybridization (FISH) is a molecular technique widely used for the detection and characterization of microbial populations. FISH is affected by a wide variety of abiotic and biotic variables and the way they interact with each other. This is translated into a wide variability of FISH procedures found in the literature. The aim of this work is to systematically study the effects of pH, dextra...


Impact of Delftia tsuruhatensis and Achromobacter xylosoxidans on Escherichia c...

Azevedo, Andreia S.; Almeida, Carina; Pereira, Bruno; Melo, Luís F.; Azevedo, Nuno F.

Recently it was demonstrated that for urinary tract infections species with a lower or unproven pathogenic potential, such as Delftia tsuruhatensis and Achromobacter xylosoxidans, might interact with conventional pathogenic agents such as Escherichia coli. Here, single- and dual-species biofilms of these microorganisms were characterized in terms of microbial composition over time, the average fitness of E. col...


FISHji: new ImageJ macros for the quantification of fluorescence in epifluoresc...

Fontenete, S.; Carvalho, D.; Lourenço, Anália; Guimarães, N.; Madureira, P.; Figueiredo, C.; Azevedo, N. F.

Fluorescence in situ hybridization (FISH) is based on the use of fluorescent staining dyes, however, the signal intensity of the images obtained by microscopy is seldom quantified with accuracy by the researcher. The development of innovative digital image processing programs and tools has been trying to overcome this problem, however, the determination of fluorescent intensity in microscopy images still has is...


Novel strategy to detect and locate periodontal pathogens: The PNA-FISH technique

Mendes, Luzia; Rocha, Rui; Azevedo, Andreia S.; Ferreira, Catarina; Henriques, Mariana; Pinto, Miguel Gonçalves; Azevedo, Nuno Filipe

Purpose: We aim to develop peptic nucleic acid (PNA) probes for the identification and localization of Aggregatibacter actinomycetemcomintans and Porphyromonas gingivalis in sub-gingival plaque and gingival biopsies by Fluorescence in situ Hybridization (FISH). Methods A PNA probe was designed for each microorganism. The PNA-FISH method was optimized to allow simultaneous hybridization of both microorganisms wi...


An in vitro model of catheter-associated urinary tract infections to investigat...

Azevedo, A. S.; Almeida, Carina; Gomes, Luciana C.; Ferreira, Carla; Mergulhão, Filipe J.; Melo, Luís F.; Azevedo, Nuno F.

Supplementary data associated with this article can be found, in the online version, at http://dx.doi.org/10.1016/j.bej.2016.11.013.; Uncommon bacteria, such as Delftia tusurhatensis have been isolated from CAUTIs in combination with well-established pathogenic bacteria such as Escherichia coli. Nonetheless, the reason why E. coli coexists with other bacteria instead of outcompeting and completely eliminating t...

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