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Detection of Salmonella enterica serovar Enteritidis using real time PCR, immun...

Almeida, Carina; Cerqueira, L.; Azevedo, N. F.; Vieira, M. J.

Several methods for the rapid and specific detection of Salmonella in food samples have been described. Here, we compare 4 of those methods in terms of assay time, procedure complexity, detection limit, sensitivity, specificity and accuracy. Milk, eggs and mayonnaise samples were artificially contaminated with Salmonella enterica serovar Enteritidis cell concentrations ranging from 1 × 10- 2 to 1 × 102 CFU per ...


Fluorescence in situ hybridization method using a peptide nucleic acid probe fo...

Machado, António; Almeida, Carina; Carvalho, Ana; Boyen, Filip; Haesebrouck, Freddy; Rodrigues, L. R.; Cerca, Nuno; Azevedo, N. F.

Lactobacillus species constitute one of the dominant and beneficial bacteria in our body and are used in developed countries as a microbial adjuvant. Identification of these probiotic bacteria is traditionally performed by culture-based techniques. However, such methods are very time-consuming and can give inaccurate results, especially when Lactobacillus is present in mixed bacterial complex communities. Our s...


Fluorescence in situ hybridization method using peptide nucleic acid probes for...

Machado, António; Almeida, Carina; Salgueiro, D. A. L.; Henriques, Ana Filipa; Vaneechoutte, Mario; Haesebrouck, Freddy; Vieira, M. J.; Rodrigues, L. R.

Background: Bacterial vaginosis (BV) is a common vaginal infection occurring in women of reproductive age. It is widely accepted that the microbial switch from normal microflora to BV is characterized by a decrease in vaginal colonization by Lactobacillus species together with an increase of Gardnerella vaginalis and other anaerobes. Our goal was to develop and optimize a novel Peptide Nucleic Acid (PNA) Fluore...


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...


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...


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

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

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 ...


Probe4pylori : a new kit for the rapid detection of H. pylori and associated cl...

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

In this study an evaluation of Probe4Pylori, a previously established PNA-FISH diagnostic test for H. pylori clarithromycin resistance in paraffin embedded gastric biopsies was performed and compared against culture followed by E-test and PCR. The Probe4Pylori showed very promising results with values of sensitivity (80%) and specificity (approximately 90%) for the patients harboring clarithromycin-resistant H....


Impact of hypoxic and anaerobic environments on multidrug-resistance of emergin...

Lopes, Susana Patrícia; Azevedo, N. F.; Pereira, M. O.

OBJECTIVES: It is well-known the establishment of steep oxygen gradients in cystic fibrosis (CF) airways mucus, giving rise to hypoxic or anaerobic zones in the deeper mucus layers, where traditional and atypical bacteria may accommodate and proliferate to biofilms. This study aimed to compare the influence of low-oxygen atmospheres on biofilm growth and susceptibility profiles of the CF-atypical species Inquil...


Synergistic interactions within polymicrobial biofilms formed by atypical and c...

Lopes, Susana Patrícia; Azevedo, N. F.; Pereira, M. O.

A complex microbial ecosystem is present in Cystic fibrosis (CF)-airways, with uncommon species co-existing and establishing dynamic interactions with traditional pathogens. This study aimed to examine in vitro biofilm formation and susceptibility patterns of two CF-atypical bacteria, Inquilinus limosus (IL) and Dolosigranulum pigrum (DP), when associated to P. aeruginosa (PA), under oxygen-atmospheres resembli...


Interaction between non-disease causing microorganism and E. coli in catheter-a...

Azevedo, Andreia; Almeida, Carina; Melo, L. F.; Azevedo, N. F.

Most biofilms involved in catheter-associated urinary tract infections (CAUTIs) are polymicrobial, with disease causing (e.g. E. coli) and non-disease causing (NDC) microorganisms (Delftia tsuruhstensis, Achromobacter xylosoxidans, Burkholderia fungorum) frequently co-inhabiting the same catheter. Nevertheless, there is a lack of knowledge about the role that NDC microorganisms have on biofilm formation by E. c...


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    Financiadores do RCAAP

Fundação para a Ciência e a Tecnologia Universidade do Minho   Governo Português Ministério da Educação e Ciência Programa Operacional da Sociedade do Conhecimento União Europeia