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Evaluation of covalent organic frameworks for the low-cost, rapid detection of shiga toxin-producing escherichia coli in ready-to-eat salads

2023-05-17Journal article Restricted access
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dc.contributor.authorRibeiro, Ana Costa
dc.contributor.authorAzinheiro, Sarah 
dc.contributor.authorFernandes, Soraia P.S. 
dc.contributor.authorLamas, Alexandre 
dc.contributor.authorPrado, Marta 
dc.contributor.authorSalonen, Laura M. 
dc.contributor.authorGarrido-Maestu, Alejandro 
dc.date.accessioned2023-10-11T17:04:57Z
dc.date.available2023-10-11T17:04:57Z
dc.date.issued2023-05-17
dc.description.abstractReady-to-eat products, such as leafy greens, must be carefully controlled as they are directly consumed without any treatment to reduce the presence of potential pathogens. Food industries, especially those that process products with short shelf-life, demand rapid detection of foodborne pathogens such as Shiga Toxin-producing Escherichia coli (STEC). In this sense, molecular methods can fulfill both requirements of turnaround time and consumer safety. The most popular rapid methods are those based on real-time PCR (qPCR) however, vegetables contain inhibitory compounds that may inhibit the amplification reaction thus, there is a need for novel sample preparation protocols. In the current study, a low-cost sample treatment based on sequential filtration steps was developed. This protocol was combined with covalent organic frameworks (COFs), and compared against a chelating resin, to evaluate their performance by multiplex qPCR targeting the major virulence genes of STEC, namely stx1, stx2, and eae, along with the rfbE for the specific identification of serogroup O157 due to its particularly high incidence, and an Internal Amplification Control to assess reaction inhibition. The optimized sample treatment effectively removed vegetable qPCR inhibitory compounds, and it was possible to detect STEC in spiked ready-to-eat salad samples in one working day, roughly 5 h, with an LOD50 of 8.7 CFU/25 g with high diagnostic sensitivity and specificity. The method was also assessed in samples with cold-stressed bacteria with good results, further demonstrating its applicability. It was demonstrated for the first time that COFs are suitable for DNA extraction and purification. In addition to this, due to the tunable nature of these materials, it is envisioned that future modifications in terms of pore size or combination with magnetic materials, will allow to further improve their performance. In addition to this, the rapid and low-cost sample treatment protocol developed demonstrated suitable for the rapid screening of STEC vegetable samples.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationCosta-Ribeiro, A., Azinheiro, S., Fernandes, S. P. S., Lamas, A., Prado, M., Salonen, L. M., & Garrido-Maestu, A. (2023). Evaluation of Covalent Organic Frameworks for the low-cost, rapid detection of Shiga Toxin-producing Escherichia coli in ready-to-eat salads. Analytica Chimica Acta, 1267, 341357. https://doi.org/10.1016/j.aca.2023.341357pt_PT
dc.identifier.doi10.1016/j.aca.2023.341357pt_PT
dc.identifier.eissn1873-4324
dc.identifier.issn0003-2670
dc.identifier.urihttp://hdl.handle.net/10400.22/23677
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S0003267023005780?via%3Dihubpt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectShigapt_PT
dc.subjectToxin-producingpt_PT
dc.subjectEscherichia colipt_PT
dc.subjectMultiplex qPCRpt_PT
dc.subjectReady-to-eat saladpt_PT
dc.subjectCovalent organic frameworkspt_PT
dc.subjectRapid methodspt_PT
dc.subjectSame-day detectionpt_PT
dc.titleEvaluation of covalent organic frameworks for the low-cost, rapid detection of shiga toxin-producing escherichia coli in ready-to-eat saladspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage7pt_PT
oaire.citation.startPage1pt_PT
oaire.citation.titleAnalytica Chimica Actapt_PT
oaire.citation.volume1267pt_PT
person.familyNameRibeiro
person.givenNameAna Costa
person.identifier.ciencia-id6211-F099-C8B4
person.identifier.orcid0000-0002-6471-7217
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication5c6b9a98-c080-4b01-bf82-68e60bc31d3c
relation.isAuthorOfPublication.latestForDiscovery5c6b9a98-c080-4b01-bf82-68e60bc31d3c

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