Development of a prototype rapid method for the detection of Escherichia coli and Salmonella enterica subspecies enterica serovar Newport based on phage amplification and detection of progeny phage by Loop Mediated Isothermal Amplification

dc.contributor.advisorFarber, Jeffrey
dc.contributor.advisorGriffiths, Mansel
dc.contributor.authorCucic, Stevan
dc.date.accessioned2017-11-02T19:04:58Z
dc.date.available2017-11-02T19:04:58Z
dc.date.copyright2017-09
dc.date.created2017-10-25
dc.date.issued2017-11-02
dc.degree.departmentDepartment of Food Scienceen_US
dc.degree.grantorUniversity of Guelphen_US
dc.degree.nameMaster of Scienceen_US
dc.degree.programmeFood Scienceen_US
dc.description.abstractThe goal of this project was to develop a rapid detection method for Escherichia coli B and Salmonella enterica subspecies enterica serovar Newport. T4 was used for the detection of E. coli B. For the purpose of detection of S. Newport, phage isolation and characterization were done. Four different phage isolates were selected and characterized by host-range analysis, transmission electron microscopy, restriction endonuclease fingerprinting, and stability under different pH and temperature conditions. A broad host-range phage called vB_SN_CGG41 that was able to infect all of the Salmonella serovars against which it was screened was selected for genome sequencing. Loop mediated isothermal amplification (LAMP) primers targeting respective unique sequences in the genomes of phages T4 and CGG41 were designed and used to detect phage nucleic acid released by boiling. Two different LAMP methods were used, which were Bioluminescent Assay in Real-Time (BART)-LAMP and endpoint colourimetric detection of pyrophosphate following conventional LAMP. Phage amplification assays (PAA) were performed with E. coli B and S. Newport to allow LAMP-based detection of phage following phage-host co-incubation.en_US
dc.identifier.urihttp://hdl.handle.net/10214/11933
dc.language.isoenen_US
dc.publisherUniversity of Guelphen_US
dc.rightsAttribution-NonCommercial-NoDerivs 2.5 Canada*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/2.5/ca/*
dc.subjectBacteriophageen_US
dc.subjectphageen_US
dc.subjectSalmonellaen_US
dc.subjectE colien_US
dc.subjectEscherichia colien_US
dc.subjectrapid detectionen_US
dc.subjectloop mediated isothermal amplificationen_US
dc.subjectphage amplification assayen_US
dc.subjectphage genomicsen_US
dc.titleDevelopment of a prototype rapid method for the detection of Escherichia coli and Salmonella enterica subspecies enterica serovar Newport based on phage amplification and detection of progeny phage by Loop Mediated Isothermal Amplificationen_US
dc.typeThesisen_US

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