Development and Application of a Novel Peptide Nucleic Acid Probe for the Specific Detection of
            <i>Cronobacter</i>
            Genomospecies (
            <i>Enterobacter sakazakii</i>
            ) in Powdered Infant Formula

Almeida, Carina; Azevedo, Nuno F.; Iversen, Carol; Fanning, Séamus; Keevil, C. W.; Vieira, M. J.

doi:10.1128/aem.02470-08

Cited by 52 publications

(48 citation statements)

References 48 publications

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“…4). This procedure was able to detect PIF samples with 1 CFU per 10 g, a value similar to that found in the Cronobacter study mentioned above (1). Salmonella detection in PIF was performed in less than 12 h, which represents a time-saving of several days compared with the ISO 6579:2002 method, recommended for Salmonella detection in food and animal feeding stuffs.…”

Section: Resultssupporting

confidence: 51%

“…It was therefore decided to apply the probe to this type of sample as previously performed for E. sakazakii (Cronobacter sp.) (1).…”

Section: Resultsmentioning

confidence: 99%

“…An experiment regarding Salmonella detection in PIF was planned based on an earlier work that also applied the PNA-FISH method to detect Cronobacter (1).…”

Section: Resultsmentioning

confidence: 99%

“…The detection of Salmonella in powdered infant formula (PIF) was based on Almeida et al (1). The infant formula (NAN 1 Premium; Nestle) was reconstituted by mixing 10 g in 80 ml of sterile distilled water.…”

Section: Methodsmentioning

confidence: 99%

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Fluorescence In Situ Hybridization Method Using a Peptide Nucleic Acid Probe for Identification of Salmonella spp. in a Broad Spectrum of Samples

Almeida

Azevedo

Fernandes

et al. 2010

Appl Environ Microbiol

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A fluorescence in situ hybridization (FISH) method for the rapid detection of Salmonella spp. using a novel peptide nucleic acid (PNA) probe was developed. The probe theoretical specificity and sensitivity were both 100%. The PNA-FISH method was optimized, and laboratory testing on representative strains from the Salmonella genus subspecies and several related bacterial species confirmed the predicted theoretical values of specificity and sensitivity. The PNA-FISH method has been successfully adapted to detect cells in suspension and is hence able to be employed for the detection of this bacterium in blood, feces, water, and powdered infant formula (PIF). The blood and PIF samples were artificially contaminated with decreasing pathogen concentrations. After the use of an enrichment step, the PNA-FISH method was able to detect 1 CFU per 10 ml of blood (5 ؋ 10 9 ؎ 5 ؋ 10 8 CFU/ml after an overnight enrichment step) and also 1 CFU per 10 g of PIF (2 ؋ 10 7 ؎ 5 ؋ 10 6 CFU/ml after an 8-h enrichment step). The feces and water samples were also enriched according to the corresponding International Organization for Standardization methods, and results showed that the PNA-FISH method was able to detect Salmonella immediately after the first enrichment step was conducted. Moreover, the probe was able to discriminate the bacterium in a mixed microbial population in feces and water by counter-staining with 4,6-diamidino-2-phenylindole (DAPI). This new method is applicable to a broad spectrum of samples and takes less than 20 h to obtain a diagnosis, except for PIF samples, where the analysis takes less than 12 h. This procedure may be used for food processing and municipal water control and also in clinical settings, representing an improved alternative to culture-based techniques and to the existing Salmonella PNA probe, Sal23S10, which presents a lower specificity.

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Section: Resultssupporting

confidence: 51%

“…It was therefore decided to apply the probe to this type of sample as previously performed for E. sakazakii (Cronobacter sp.) (1).…”

Section: Resultsmentioning

confidence: 99%

“…An experiment regarding Salmonella detection in PIF was planned based on an earlier work that also applied the PNA-FISH method to detect Cronobacter (1).…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Fluorescence In Situ Hybridization Method Using a Peptide Nucleic Acid Probe for Identification of Salmonella spp. in a Broad Spectrum of Samples

Almeida

Azevedo

Fernandes

et al. 2010

Appl Environ Microbiol

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“…An important feature to bear in mind when optimizing FISH protocols is that some food components may present a strong autofluorescence signal, which can interfere with bacterial detection (32). To eliminate/reduce this phenomenon, an additional step can be added before the hybridization procedure.…”

Section: Resultsmentioning

confidence: 99%

Detection of Escherichia coli O157 by Peptide Nucleic Acid Fluorescence In Situ Hybridization (PNA-FISH) and Comparison to a Standard Culture Method

Almeida

Sousa

Rocha³

et al. 2013

Appl Environ Microbiol

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Despite the emergence of non-O157 Shiga toxin-producing Escherichia coli (STEC) infections, E. coli serotype O157 is still the most commonly identified STEC in the 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 the gold standard. Here, we propose a new peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) method for the rapid detection of E. coli O157. Testing on 54 representative strains showed that the PNA probe is highly sensitive and specific to E. coli O157. The method then was optimized for detection in food samples. Ground beef and unpasteurized milk samples were artificially contaminated with E. coli O157 concentrations ranging from 1 ؋ 10 ؊2 to 1 ؋

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Cronobacterspp.

Yan¹,

Power²,

Tall³

et al. 2013

Guide to Foodborne Pathogens

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Development and Application of a Novel Peptide Nucleic Acid Probe for the Specific Detection of Cronobacter Genomospecies ( Enterobacter sakazakii ) in Powdered Infant Formula

Cited by 52 publications

References 48 publications

Fluorescence In Situ Hybridization Method Using a Peptide Nucleic Acid Probe for Identification of Salmonella spp. in a Broad Spectrum of Samples

Fluorescence In Situ Hybridization Method Using a Peptide Nucleic Acid Probe for Identification of Salmonella spp. in a Broad Spectrum of Samples

Detection of Escherichia coli O157 by Peptide Nucleic Acid Fluorescence In Situ Hybridization (PNA-FISH) and Comparison to a Standard Culture Method

Cronobacterspp.

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