Design of DNA microarrays for pathogen detection

A method using DNA Microarrays following PCR amplification of 12 selected unique genes has been proposed for the specific detection of Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Typhi, Salmonella Enteritidis and Yersinia Enterocolitica in drinking water sample. The use of specific...

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Bibliographic Details
Main Author: Lau, Yi Xiang.
Other Authors: Chang Wook, Matthew
Format: Final Year Project (FYP)
Language:English
Published: 2010
Subjects:
Online Access:http://hdl.handle.net/10356/39369
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author Lau, Yi Xiang.
author2 Chang Wook, Matthew
author_facet Chang Wook, Matthew
Lau, Yi Xiang.
author_sort Lau, Yi Xiang.
collection NTU
description A method using DNA Microarrays following PCR amplification of 12 selected unique genes has been proposed for the specific detection of Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Typhi, Salmonella Enteritidis and Yersinia Enterocolitica in drinking water sample. The use of specific literature PCR primers amplifying the unique rfbE sequence as well as hybridization of amplicons with specific DNA Microarray probes was proven to be able to provide a specific detection of E. coli O157:H7 in theory. 3 other genes, with primers and DNA probes derived from these genes (fliC, stx1 and stx2) were used as confirmation to detection results. Moreover, the STM4497 gene was unique and its primer set was also proven to be specific to S. Typhimurium. Also, since the STM4497 Microarray probe was specific for the same pathogen, it should provide a specific detection of S. Typhimurium, using the STM4492 primer sets and Microarray probe as confirmation. In addition, it was also proven theoretically that the use of the viaB primer set to amplify the unique viaB gene sequence allowed the specific detection of Salmonella Typhi using specific DNA Microarray probes. Tyv PCR primer set and its DNA Microarray probe provided a good confirmation of the presence of S. Typhi. Also, the amplification of the unique gene of S. Enteritidis – the spvA gene, using specific PCR primer set was able to allow the specific detection of S. Enteritidis by DNA Microarray. Finally, the ail and yst genes were proven to be unique to Y. Enterocolitica. The PCR primer sets used were proven to be specific and their use provided a theoretically specific detection of Y. Enterocolitica through hybridization with DNA Microarray probes. The virF primers and DNA Microarray probes are expected to give a good confirmation of the hybridization results.
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spelling ntu-10356/393692023-03-03T15:40:51Z Design of DNA microarrays for pathogen detection Lau, Yi Xiang. Chang Wook, Matthew School of Chemical and Biomedical Engineering DRNTU::Engineering::Chemical engineering::Biotechnology A method using DNA Microarrays following PCR amplification of 12 selected unique genes has been proposed for the specific detection of Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Typhi, Salmonella Enteritidis and Yersinia Enterocolitica in drinking water sample. The use of specific literature PCR primers amplifying the unique rfbE sequence as well as hybridization of amplicons with specific DNA Microarray probes was proven to be able to provide a specific detection of E. coli O157:H7 in theory. 3 other genes, with primers and DNA probes derived from these genes (fliC, stx1 and stx2) were used as confirmation to detection results. Moreover, the STM4497 gene was unique and its primer set was also proven to be specific to S. Typhimurium. Also, since the STM4497 Microarray probe was specific for the same pathogen, it should provide a specific detection of S. Typhimurium, using the STM4492 primer sets and Microarray probe as confirmation. In addition, it was also proven theoretically that the use of the viaB primer set to amplify the unique viaB gene sequence allowed the specific detection of Salmonella Typhi using specific DNA Microarray probes. Tyv PCR primer set and its DNA Microarray probe provided a good confirmation of the presence of S. Typhi. Also, the amplification of the unique gene of S. Enteritidis – the spvA gene, using specific PCR primer set was able to allow the specific detection of S. Enteritidis by DNA Microarray. Finally, the ail and yst genes were proven to be unique to Y. Enterocolitica. The PCR primer sets used were proven to be specific and their use provided a theoretically specific detection of Y. Enterocolitica through hybridization with DNA Microarray probes. The virF primers and DNA Microarray probes are expected to give a good confirmation of the hybridization results. Bachelor of Engineering (Chemical and Biomolecular Engineering) 2010-05-21T06:23:39Z 2010-05-21T06:23:39Z 2010 2010 Final Year Project (FYP) http://hdl.handle.net/10356/39369 en Nanyang Technological University 72 p. application/pdf
spellingShingle DRNTU::Engineering::Chemical engineering::Biotechnology
Lau, Yi Xiang.
Design of DNA microarrays for pathogen detection
title Design of DNA microarrays for pathogen detection
title_full Design of DNA microarrays for pathogen detection
title_fullStr Design of DNA microarrays for pathogen detection
title_full_unstemmed Design of DNA microarrays for pathogen detection
title_short Design of DNA microarrays for pathogen detection
title_sort design of dna microarrays for pathogen detection
topic DRNTU::Engineering::Chemical engineering::Biotechnology
url http://hdl.handle.net/10356/39369
work_keys_str_mv AT lauyixiang designofdnamicroarraysforpathogendetection