Development And Evaluation Of Novel PCR Assays For The Sensitive, Rapid And Accurate Detection Of Salmonella Spp., L. Monocytogenes, E. Coli O157:H7 And G. Stearothermofillus In Pasteurized Milk And Fresh Juice Products | 17166
Journal of Biotechnology & Biomaterials
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Development and evaluation of novel PCR assays for the sensitive, rapid and accurate detection of Salmonella spp., L. monocytogenes, E. coli O157:H7 and G. stearothermofillus in pasteurized milk and fresh juice products
5th World Congress on Biotechnology
A Scorilas, A Kladi-Skandali, D Ghikas, F Gaitis, E Beletsiotis, K Kalantzi and M Avgeris
Foodborne illnesses are related with high rates of morbidity and mortality as well as with substantial economic burden.
The replacement of traditional and laborious protocols by sensitive and rapid methods is required, and PCR-based
assays represent the corner stone of this effort. A multiplex PCR assay was developed for the
Salmonella spp., Listeria monocytogenes
O157:H7, using specific primers which target
respectively. Quantitative TaqMan-based Real-time PCR (qPCR) methodology was developed for their quantification as well
. Pasteurized milk and fresh orange juice were spiked and cultured based to ISO methods.
Samples were taken for DNA extraction and PCR analysis every two hours until 8 hours of culture. Assay specificity was
individually assessed, using DNA from closely related species. Concerning singleplex qPCR, the detection limits for
were 5x10-4 ng DNA, while for
Using multiplex PCR assay, detection of the pathogens was succeeded following 4 hours of culture spiked with 10 cfu/ml or
more, while accurate quantification of the pathogens was also performed for the same samples and all the spiked pathogens. We
developed two specific and sensitive PCR assays for the simultaneous detection of three different food-associated pathogens,
paving the way for their application in food analysis.
The present work was funded by SYNERGASIA 2009 PROGRAMME. This Programme is co-funded by
the European Regional Development Fund and National Resources. (Project code: 09SYN-22-977). Partial expenses for the
presentation of this abstract were funded by the University of Athens Special Account of Research Grants no 10812
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