Development of a targeted proteomic assay for rapid detection of Shiga-like toxins 1 and 2 in Shiga toxin-producing Escherichia coli

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dc.contributor.authorScharikow, Leanne Gene
dc.contributor.examiningcommitteeMcClarty, Grant (Medical Microbiology and Infectious Diseases) Nadon, Celine (Medical Microbiology and Infectious Diseases) Narvaez, Claudia (Food Science)en_US
dc.contributor.supervisorGraham, Morag (Medical Microbiology and Infectious Diseases)en_US
dc.date.accessioned2017-01-05T21:33:21Z
dc.date.available2017-01-05T21:33:21Z
dc.date.issued2016
dc.degree.disciplineMedical Microbiologyen_US
dc.degree.levelMaster of Science (M.Sc.)en_US
dc.description.abstractShiga toxin-producing Escherichia coli (STEC) are extensive contributors to foodborne illness, causing renal and central nervous system damage due to production of Shiga toxin (Stx). Rapid Stx detection is important to distinguish STEC from other enteric pathogens. Current detection techniques are time consuming, expensive, and lack sensitivity. We have developed and evaluated a novel targeted mass spectrometry-based assay for detection of Stx using parallel reaction monitoring (PRM). The PRM assay used 11 target tryptic peptides and was validated using STEC and non-STEC bacterial cultures. Stx was detected in 56 of 62 STEC isolates and did not detect Stx in any of the 29 non-STEC isolates. The PRM assay successfully determined the Stx2 subtype in 32 of 46 Stx2-positive isolates. By applying a targeted proteomics assay, we were able to simultaneously detect Stx toxins 1 and 2 and subtype Stx2 into six toxin subgroups in Stx2-positive isolates.en_US
dc.description.noteFebruary 2017en_US
dc.identifier.urihttp://hdl.handle.net/1993/31991
dc.language.isoengen_US
dc.rightsopen accessen_US
dc.subjectMass spectrometryen_US
dc.subjectTargeted proteomicsen_US
dc.subjectShiga toxin-producing Escherichia colien_US
dc.subjectShiga toxinen_US
dc.titleDevelopment of a targeted proteomic assay for rapid detection of Shiga-like toxins 1 and 2 in Shiga toxin-producing Escherichia colien_US
dc.typemaster thesisen_US
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