Salmonella Detection in Food Using a HEK-hTLR5 Reporter Cell-Based Sensor

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dc.authoridEser, Esma / 0000-0002-9681-8442
dc.contributor.authorEser, Esma
dc.contributor.authorFelton, Victoria A.
dc.contributor.authorDrolia, Rishi
dc.contributor.authorBhunia, Arun K.
dc.date.accessioned2025-01-27T20:49:36Z
dc.date.available2025-01-27T20:49:36Z
dc.date.issued2024
dc.departmentÇanakkale Onsekiz Mart Üniversitesi
dc.description.abstractThe development of a rapid, sensitive, specific method for detecting foodborne pathogens is paramount for supplying safe food to enhance public health safety. Despite the significant improvement in pathogen detection methods, key issues are still associated with rapid methods, such as distinguishing living cells from dead, the pathogenic potential or health risk of the analyte at the time of consumption, the detection limit, and the sample-to-result. Mammalian cell-based assays analyze pathogens' interaction with host cells and are responsive only to live pathogens or active toxins. In this study, a human embryonic kidney (HEK293) cell line expressing Toll-Like Receptor 5 (TLR-5) and chromogenic reporter system (HEK dual hTLR5) was used for the detection of viable Salmonella in a 96-well tissue culture plate. This cell line responds to low concentrations of TLR5 agonist flagellin. Stimulation of TLR5 ligand activates nuclear factor-kB (NF-kappa B)-linked alkaline phosphatase (AP-1) signaling cascade inducing the production of secreted embryonic alkaline phosphatase (SEAP). With the addition of a rho-nitrophenyl phosphate as a substrate, a colored end product representing a positive signal is quantified. The assay's specificity was validated with the top 20 Salmonella enterica serovars and 19 non-Salmonella spp. The performance of the assay was also validated with spiked food samples. The total detection time (sample-to-result), including shortened pre-enrichment (4 h) and selective enrichment (4 h) steps with artificially inoculated outbreak-implicated food samples (chicken, peanut kernel, peanut butter, black pepper, mayonnaise, and peach), was 15 h when inoculated at 1-100 CFU/25 g sample. These results show the potential of HEK-Dual (TM) hTLR5 cell-based functional biosensors for the rapid screening of Salmonella.
dc.description.sponsorshipScientific and Technological Research Council of Turkey [1059B191800917]; Scientific and Technological Research Council of Turkey (TUBITAK)-BIDEB [1016249]; National Institute of Food and Agriculture of USDA [2023-67017-40051]; USDA-NIFA
dc.description.sponsorshipThis research was funded by the Scientific and Technological Research Council of Turkey (TUBITAK)-BIDEB under the 2219/International Postdoctoral Research Fellowship Programme with Application Number 1059B191800917. The research was supported by funds from the National Institute of Food and Agriculture of USDA (Hatch grant # 1016249; A.K.B.), the start-up funds at ODU (R.D.), and USDA-NIFA (Award # 2023-67017-40051; R.D.).
dc.identifier.doi10.3390/bios14090444
dc.identifier.issn2079-6374
dc.identifier.issue9
dc.identifier.pmid39329819
dc.identifier.scopus2-s2.0-85205151826
dc.identifier.scopusqualityQ1
dc.identifier.urihttps://doi.org/10.3390/bios14090444
dc.identifier.urihttps://hdl.handle.net/20.500.12428/25245
dc.identifier.volume14
dc.identifier.wosWOS:001326363200001
dc.identifier.wosqualityN/A
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.language.isoen
dc.publisherMdpi
dc.relation.ispartofBiosensors-Basel
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/openAccess
dc.snmzKA_WoS_20250125
dc.subjectSalmonella
dc.subjectcell-based sensor
dc.subjectHEK-dual hTLR 5
dc.subjectdetection
dc.subjectflagella
dc.subjectimmunomagnetic separation
dc.subjectfood safety
dc.titleSalmonella Detection in Food Using a HEK-hTLR5 Reporter Cell-Based Sensor
dc.typeArticle

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