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Detection of Salmonella enterica serovar Enteritidis using real time PCR, immunocapture assay, PNA FISH and standard culture methods in different types of food samples.
MedLine Citation:
PMID:  23246608     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
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 1×10(2)CFU per 25g or ml of food. Samples were then pre-enriched and analyzed by either: i) real-time PCR, using the iQ-Check Salmonella kit; ii) immunocapture, using the RapidChek SELECT Salmonella; iii) a peptide nucleic acid fluorescence in situ hybridization (PNA FISH) method and iv) the traditional bacteriological method ISO 6579:2002. All methods were able to detect Salmonella in the different types of food matrixes and presented a similar detection level of 1CFU per 25g or ml of food sample. The immunocapture and the PNA FISH methods proved to be very reliable, as their results were 100% in agreement with the ISO method. However, real-time PCR presented a significant number of false positives, which resulted in a specificity of 55.6% (CI 95%, 31.3-77.6) and an accuracy of 82.2% (CI 95%, 63.2-91.4) for this method. Sensitivity was 100% since no false negative results were observed. In conclusion, the implementation of these molecular techniques, mainly the immunocapture and PNA-FISH methods, provides a reliable and less time-consuming alternative for the detection of Salmonella spp. in food samples.
Authors:
C Almeida; L Cerqueira; N F Azevedo; M J Vieira
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-11-23
Journal Detail:
Title:  International journal of food microbiology     Volume:  161     ISSN:  1879-3460     ISO Abbreviation:  Int. J. Food Microbiol.     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-12-18     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8412849     Medline TA:  Int J Food Microbiol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  16-22     Citation Subset:  -    
Copyright Information:
Copyright © 2012 Elsevier B.V. All rights reserved.
Affiliation:
IBB, Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar 4710-057, Braga, Portugal; LEPAE, Department of Chemical Engineering, Faculty of Engineering, University of Porto, Porto, Portugal; BioMode, Biomolecular Determination S.A., SpinPark - Centro de Incubação de Base Tecnológica, Avepark - Zona Industrial da Gandra, 4806-909 Caldas das Taipas, Guimarães, Portugal.
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