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Permanent link (DOI): https://doi.org/10.7939/R3W08WP6K

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Detection of Shiga Toxin-Producing Escherichia coli Using Loop Mediated Isothermal Amplification Open Access

Descriptions

Other title
Subject/Keyword
optimization
detection
shiga toxin
escherichia coli
loop-mediated isothermal amplification
nucleic acid amplification
verotoxin
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
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.
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File format: pdf (Portable Document Format)
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File size: 9834929
Last modified: 2015:10:12 20:15:41-06:00
Filename: Allen_Richelle_Fall2013.pdf
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Status message: File header gives version as 1.4, but catalog dictionary gives version as 1.3
File title: Microsoft Word - STUPIDTHESIS.docx
File author: Richelle Allen
Page count: 105
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