Optimization of multiplex PCR conditions for rapid detection of Escherichia coli O157:H7 virulence genes
The main source of E. coli 0157:H7 is cattle, but recent studies showed high percentage of outbreaks contributed by contaminated water. The occurrence of E. coli O157:H7 in environmental water samples poses a potential threat to human health. The aim of this study was to establish a protocol for the...
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| Format: | Article |
| Language: | English |
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Faculty of Food Science and Technology, Universiti Putra Malaysia
2012
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| Online Access: | http://psasir.upm.edu.my/id/eprint/41042/ http://psasir.upm.edu.my/id/eprint/41042/1/Optimization%20of%20multiplex%20PCR%20conditions%20for%20rapid%20detection%20of%20Escherichia%20coli%20O157H7%20virulence%20genes.pdf |
| _version_ | 1848849589605498880 |
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| author | Ranjit Singh, Jeshveen Singh Chai, Lay Ching Pui, Chai Fung Radu, Son |
| author_facet | Ranjit Singh, Jeshveen Singh Chai, Lay Ching Pui, Chai Fung Radu, Son |
| author_sort | Ranjit Singh, Jeshveen Singh |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | The main source of E. coli 0157:H7 is cattle, but recent studies showed high percentage of outbreaks contributed by contaminated water. The occurrence of E. coli O157:H7 in environmental water samples poses a potential threat to human health. The aim of this study was to establish a protocol for the detection of the pathogen E. coli O157:H7 and E. coli virulence genes (eaeA, rfbE, hly, stx 1, and stx 2) in a multiplex PCR protocol using six specific primer pairs. The target genes produced species-specific amplicons at 625 bp, 397 bp, 296 bp, 166 bp, 210 bp and 484 bp for E. coli O157:H7 (fliC h7 gene) and virulence genes (eaeA, rfbE, hly, stx 1, and stx 2) respectively. The results obtained show that the established PCR protocol is suitable for a rapid and specific analysis of the pathogenic E. coli O157:H7 in environmental water samples for the assessment of microbiological risks. |
| first_indexed | 2025-11-15T09:52:48Z |
| format | Article |
| id | upm-41042 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T09:52:48Z |
| publishDate | 2012 |
| publisher | Faculty of Food Science and Technology, Universiti Putra Malaysia |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-410422020-07-14T04:46:17Z http://psasir.upm.edu.my/id/eprint/41042/ Optimization of multiplex PCR conditions for rapid detection of Escherichia coli O157:H7 virulence genes Ranjit Singh, Jeshveen Singh Chai, Lay Ching Pui, Chai Fung Radu, Son The main source of E. coli 0157:H7 is cattle, but recent studies showed high percentage of outbreaks contributed by contaminated water. The occurrence of E. coli O157:H7 in environmental water samples poses a potential threat to human health. The aim of this study was to establish a protocol for the detection of the pathogen E. coli O157:H7 and E. coli virulence genes (eaeA, rfbE, hly, stx 1, and stx 2) in a multiplex PCR protocol using six specific primer pairs. The target genes produced species-specific amplicons at 625 bp, 397 bp, 296 bp, 166 bp, 210 bp and 484 bp for E. coli O157:H7 (fliC h7 gene) and virulence genes (eaeA, rfbE, hly, stx 1, and stx 2) respectively. The results obtained show that the established PCR protocol is suitable for a rapid and specific analysis of the pathogenic E. coli O157:H7 in environmental water samples for the assessment of microbiological risks. Faculty of Food Science and Technology, Universiti Putra Malaysia 2012 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/41042/1/Optimization%20of%20multiplex%20PCR%20conditions%20for%20rapid%20detection%20of%20Escherichia%20coli%20O157H7%20virulence%20genes.pdf Ranjit Singh, Jeshveen Singh and Chai, Lay Ching and Pui, Chai Fung and Radu, Son (2012) Optimization of multiplex PCR conditions for rapid detection of Escherichia coli O157:H7 virulence genes. International Food Research Journal, 19 (2). pp. 461-466. ISSN 1985-4668; ESSN: 2231-7546 http://www.ifrj.upm.edu.my/19%20%2802%29%202012/%2812%29IFRJ-2012%20Jeshveen.pdf |
| spellingShingle | Ranjit Singh, Jeshveen Singh Chai, Lay Ching Pui, Chai Fung Radu, Son Optimization of multiplex PCR conditions for rapid detection of Escherichia coli O157:H7 virulence genes |
| title | Optimization of multiplex PCR conditions for rapid detection of Escherichia coli O157:H7 virulence genes |
| title_full | Optimization of multiplex PCR conditions for rapid detection of Escherichia coli O157:H7 virulence genes |
| title_fullStr | Optimization of multiplex PCR conditions for rapid detection of Escherichia coli O157:H7 virulence genes |
| title_full_unstemmed | Optimization of multiplex PCR conditions for rapid detection of Escherichia coli O157:H7 virulence genes |
| title_short | Optimization of multiplex PCR conditions for rapid detection of Escherichia coli O157:H7 virulence genes |
| title_sort | optimization of multiplex pcr conditions for rapid detection of escherichia coli o157:h7 virulence genes |
| url | http://psasir.upm.edu.my/id/eprint/41042/ http://psasir.upm.edu.my/id/eprint/41042/ http://psasir.upm.edu.my/id/eprint/41042/1/Optimization%20of%20multiplex%20PCR%20conditions%20for%20rapid%20detection%20of%20Escherichia%20coli%20O157H7%20virulence%20genes.pdf |