Microelectronic-sensing assay to detect presence of Verotoxins in human faecal samples

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DOIResolve DOI: http://doi.org/10.1111/j.1365-2672.2012.05321.x
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TypeArticle
Journal titleJournal of Applied Microbiology
ISSN1364-5072
Volume113
Issue2
Pages429437; # of pages: 9
SubjectEscherichia coli; real-time cell electronic sensing assay; vero cell assay; verotoxin assay
AbstractAims: To develop a novel Vero cell assay that implements a real-time cell electronic sensing (RT-CES) system for the determination of the presence of verotoxin-producing Escherichia coli (VTEC). The assay overcomes the major drawbacks in conventional Vero cell assay, for example, labour-intensive and time-consuming. Methods and Results: Cells were grown onto the surfaces of microelectronic sensors that are integrated into the bottom surfaces of the microtiter plate. Cellular viability was monitored in real-time and quantified based on changes in the sensor's electrical impedance. For cell viability measurement, the data generated on the RT-CES system correlated well with those obtained by the Vero cell assay for Verotoxins. To assess cytotoxicity, test cells growing on microelectronic sensors were treated with either supernatant from pure cultures, or stool samples. The specific neutralizing antibodies of VT1 and VT2 were used to identify specific toxins in the samples. Conclusions: The RT-CES assay provides a sensitive measurement comparable to conventional crystal violet assay. The assay has been successfully and specifically used to identify VTEC in human faecal samples. Significance and Impact of the Study: The RT-CES assay significantly shortens the testing time from 48 to 72h required by the crystal violet assay to only 15h with automated operation.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC); Security and Disruptive Technologies
Peer reviewedYes
NPARC number21270226
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Record identifierd5c0741a-0f7f-4d01-8935-2f5dd4920a29
Record created2014-01-14
Record modified2016-05-09
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