Detection of Shiga Toxin-Producing Escherichia coli Using Loop Mediated Isothermal AmplificationReport as inadecuate




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optimization, detection, shiga toxin, escherichia coli, loop-mediated isothermal amplification, nucleic acid amplification, verotoxin

Allen,Richelle J

Supervisor and department: Chui,Linda Laboratory Medicine and Pathology Tyrrell,Greg Laboratory Medicine and Pathology

Examining committee member and department: Chui,Linda Laboratory Medicine and Pathology Tyrrell,Greg Laboratory Medicine and Pathology Keelan,Monika Laboratory Medicine and Pathology McMullen,Lynn Agriculture, Life and Environmental Sciences

Department: Medical Sciences- Laboratory Medicine and Pathology

Specialization:

Date accepted: 2013-08-29T09:40:41Z

Graduation date: 2013-11

Degree: Master of Science

Degree level: Master's

Abstract: Shiga toxin-producing Escherichia coli STEC is a worldwide health concern. To detect STEC, a loop mediated isothermal amplification reaction LAMP was optimized to detect shiga toxin genes. LAMP’s performance was compared to conventional and real-time RT PCR using two product detection methods. All three DNA amplification methods produced similar results. LAMP performed well when tested with randomly selected stool samples. LAMP, with agarose gel detection, showed a sensitivity of 90%-100%, specificity of 95%-99%, a positive predictive value of 56%-89% and a negative predictive value of 99%-100% for stx1 and stx2 respectively. Sybr Green 1 detection had a sensitivity of 100%-100%, specificity of 93%-77%, positive predictive values of 76-39% and negative predictive values of 100-100% for stx1 and stx2 respectively. Per 10 tests LAMP costs approximately $45, when using a rapid lysis DNA extraction and agarose gel electrophoresis product detection. LAMP could be implemented in laboratories without dedicated molecular biology facilities.

Language: English

DOI: doi:10.7939-R3W08WP6K

Rights: Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.





Author: Allen,Richelle J

Source: https://era.library.ualberta.ca/



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