A Rapid and Cost-Efficient Technique for Simultaneous/Duplex Detection of Listeria Monocytogenes and Escherichia Coli O157:H7 Using Real Time PCR

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dc.authoridKaynak, Ahmet/0000-0001-6551-1125
dc.contributor.authorKaynak, Ahmet
dc.contributor.authorSakalar, Ergun
dc.date.accessioned2025-01-27T21:03:34Z
dc.date.available2025-01-27T21:03:34Z
dc.date.issued2016
dc.departmentÇanakkale Onsekiz Mart Üniversitesi
dc.description.abstractThe increasing industrial interest in easy, economical and reliable methods has led to the development and application of DNA-based methods for the detection of microbial pathogens in food. In the present article, we describe the development of a cost-efficient EvaGreen-based real-time Polymerase chain reaction (PCR) technique for simultaneous and practical detection of Listeria monocytogenes (L. monocytogenes) and Escherichia coli O157:H7 (E. coli O157:H7) in the food matrix. EvaGreen-based simplex and duplex real-time PCR showed that specific PCR products were identified by melting curve analysis, and had a reproducible T-m of 77.000.4C for L. monocytogenes and 85.60 +/- 0.2 for E. coli O157:H7.The absolute detection limit of Eva Green-based simplex and duplex real-time PCR was 0.0001 ng/L for DNA of both pathogens. The relative detection limit of the simplex and duplex technique was less than 10 cells/mL for the two pathogens. Practical ApplicationsThis study illustrates a rapid, effective and cheap identification method to simultaneously monitor two foodborne pathogens in food samples. The specificity and sensitivity of this method can be used to clearly identify these two foodborne pathogens in practical food samples based on the specific genes of the species. Hence, this detection method is applicable to surveillance measures for these two foodborne pathogens in the food production chain. Moreover, this technique is used by food control laboratories in a secure way.
dc.description.sponsorshipScientific Research Project Office (SRPO) of Canakkale Onsekiz Mart University [FYL-2014-255]; Scientific and Technological Research Council of Turkey (TUBITAK) [113Z831]
dc.description.sponsorshipThis research was supported by Scientific Research Project Office (SRPO) of Canakkale Onsekiz Mart University with Project Grant No. FYL-2014-255. The authors would like to thank SRPO. The authors would also like to thank Food Gene Analytic DNA Technology Designing (GIDAGEN), TETRAT Biotechnology Company and The Scientific and Technological Research Council of Turkey (TUBITAK) (Project Grant No. 113Z831) that they provided necessary laboratory facilities.
dc.identifier.doi10.1111/jfs.12254
dc.identifier.endpage382
dc.identifier.issn0149-6085
dc.identifier.issn1745-4565
dc.identifier.issue3
dc.identifier.scopus2-s2.0-84954286956
dc.identifier.scopusqualityQ2
dc.identifier.startpage375
dc.identifier.urihttps://doi.org/10.1111/jfs.12254
dc.identifier.urihttps://hdl.handle.net/20.500.12428/27338
dc.identifier.volume36
dc.identifier.wosWOS:000380939600011
dc.identifier.wosqualityQ3
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.language.isoen
dc.publisherWiley
dc.relation.ispartofJournal of Food Safety
dc.relation.publicationcategoryinfo:eu-repo/semantics/openAccess
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.snmzKA_WoS_20250125
dc.subjectToxin Genes Stx1
dc.subjectSalmonella Spp.
dc.subjectMultiplex Pcr
dc.subjectMeat-Products
dc.subjectMilk
dc.subjectO157-H7
dc.subjectVegetables
dc.subjectO157/H7
dc.subjectSamples
dc.subjectAssays
dc.titleA Rapid and Cost-Efficient Technique for Simultaneous/Duplex Detection of Listeria Monocytogenes and Escherichia Coli O157:H7 Using Real Time PCR
dc.typeArticle

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