Multiplex PCR for Simultaneous Detection of Virulence Genes in Escherichia coli
Diarrhea caused by Escherichia coli infection is a major cause of public health problems in developing countries. In view of the deficits and limitations of conventional methods of detecting the virulence determinants, a multiplex Polymerase Chain Reaction (PCR) assay was optimized and developed to...
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Faculty of Science, University of Malaya
2009
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| Online Access: | http://eprints.um.edu.my/5657/1/Multiplex_PCR_for_simultaneous_detection_of_virulence_Genes_in_Escherichia_coli.pdf http://eprints.um.edu.my/5657/ https://mjs.um.edu.my/index.php/MJS/article/view/7682/5277 |
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| author | Thong, Kwai Lin Yu, Ke Xin |
| author_facet | Thong, Kwai Lin Yu, Ke Xin |
| author_sort | Thong, Kwai Lin |
| building | UM Library |
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| content_provider | Universiti Malaya |
| content_source | UM Research Repository |
| continent | Asia |
| country | Malaysia |
| description | Diarrhea caused by Escherichia coli infection is a major cause of public health problems in developing countries. In view of the deficits and limitations of conventional methods of detecting the virulence determinants, a multiplex Polymerase Chain Reaction (PCR) assay was optimized and developed to provide an effective, rapid and specific diagnostic tool to simultaneously detect virulence genes such as heatstable toxin 1 (ST1), heat-labile toxin 1 (LT1), heat-labile toxin 2 (LT2), verotoxin1 (VT1), verotoxin 2 (VT2) and attachment and effacement (eaeA) in pathogenic E. coli. Five sets of primers targeting these six virulence genes were optimized by using postitive control E. coli strains. The optimized conditions consisted of 3.0 mM of MgCl2, 0.2 mM of dNTPs, 1.5 U of Taq DNA polymerase (Promega), 0.70 µM of VT primer, 0.60 µM of LT2 primer and 0.07 µM each of LT1 primer, ST primer and AE primer. The mPCR assay was then applied to a panel of 87 E. coli isolates from different sources. One food isolate (EC 375) was positive for eaeA gene while another environmental isolate had ST, LT1, eaeA and VT genes. The study shows that the mPCR assay is a useful tool to differentiate the pathogenic potential (pathotypes) of E. coli by presence of known virulence genes. |
| format | Article |
| id | my.um.eprints-5657 |
| institution | Universiti Malaya |
| language | en |
| publishDate | 2009 |
| publisher | Faculty of Science, University of Malaya |
| record_format | eprints |
| spelling | my.um.eprints-56572018-10-15T03:44:27Z http://eprints.um.edu.my/5657/ Multiplex PCR for Simultaneous Detection of Virulence Genes in Escherichia coli Thong, Kwai Lin Yu, Ke Xin Q Science (General) QH Natural history QR Microbiology Diarrhea caused by Escherichia coli infection is a major cause of public health problems in developing countries. In view of the deficits and limitations of conventional methods of detecting the virulence determinants, a multiplex Polymerase Chain Reaction (PCR) assay was optimized and developed to provide an effective, rapid and specific diagnostic tool to simultaneously detect virulence genes such as heatstable toxin 1 (ST1), heat-labile toxin 1 (LT1), heat-labile toxin 2 (LT2), verotoxin1 (VT1), verotoxin 2 (VT2) and attachment and effacement (eaeA) in pathogenic E. coli. Five sets of primers targeting these six virulence genes were optimized by using postitive control E. coli strains. The optimized conditions consisted of 3.0 mM of MgCl2, 0.2 mM of dNTPs, 1.5 U of Taq DNA polymerase (Promega), 0.70 µM of VT primer, 0.60 µM of LT2 primer and 0.07 µM each of LT1 primer, ST primer and AE primer. The mPCR assay was then applied to a panel of 87 E. coli isolates from different sources. One food isolate (EC 375) was positive for eaeA gene while another environmental isolate had ST, LT1, eaeA and VT genes. The study shows that the mPCR assay is a useful tool to differentiate the pathogenic potential (pathotypes) of E. coli by presence of known virulence genes. Faculty of Science, University of Malaya 2009 Article PeerReviewed application/pdf en http://eprints.um.edu.my/5657/1/Multiplex_PCR_for_simultaneous_detection_of_virulence_Genes_in_Escherichia_coli.pdf Thong, Kwai Lin and Yu, Ke Xin (2009) Multiplex PCR for Simultaneous Detection of Virulence Genes in Escherichia coli. Malaysian Journal of Science, 28 (1). pp. 1-14. ISSN 1394-3065, DOI https://doi.org/10.22452/mjs.vol28no1.1 <https://doi.org/10.22452/mjs.vol28no1.1>. https://mjs.um.edu.my/index.php/MJS/article/view/7682/5277 doi:10.22452/mjs.vol28no1.1 |
| spellingShingle | Q Science (General) QH Natural history QR Microbiology Thong, Kwai Lin Yu, Ke Xin Multiplex PCR for Simultaneous Detection of Virulence Genes in Escherichia coli |
| title | Multiplex PCR for Simultaneous Detection of Virulence Genes in Escherichia coli |
| title_full | Multiplex PCR for Simultaneous Detection of Virulence Genes in Escherichia coli |
| title_fullStr | Multiplex PCR for Simultaneous Detection of Virulence Genes in Escherichia coli |
| title_full_unstemmed | Multiplex PCR for Simultaneous Detection of Virulence Genes in Escherichia coli |
| title_short | Multiplex PCR for Simultaneous Detection of Virulence Genes in Escherichia coli |
| title_sort | multiplex pcr for simultaneous detection of virulence genes in escherichia coli |
| topic | Q Science (General) QH Natural history QR Microbiology |
| url | http://eprints.um.edu.my/5657/1/Multiplex_PCR_for_simultaneous_detection_of_virulence_Genes_in_Escherichia_coli.pdf http://eprints.um.edu.my/5657/ https://mjs.um.edu.my/index.php/MJS/article/view/7682/5277 |
| url_provider | http://eprints.um.edu.my/ |