Real-time PCR for detection of fliC gene of E. coli O157:H7 in beef and chicken meat
The SYBR Green I real-time PCR assay was used to quantify E. coli O157:H7 in various meat samples. Primers were designed to amplify and quantify the structural flagella (fliC) gene of E. coli O157:H7 in a single reaction. The primer specificity was confirmed with DNA from an ATCC culture of E. coli...
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| Format: | Article |
| Language: | English |
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Malaysian Agricultural Research and Development Institute (MARDI)
2012
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| Online Access: | http://psasir.upm.edu.my/id/eprint/49301/1/Real-time%20PCR%20for%20detection%20of%20fliC%20gene%20of%20E.%20coli%20O157H7%20in%20beef%20and%20chicken%20meat.pdf |
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| author | Mohd Saad, Suria Abdul Kadir, Adlin Azlina Ishak, Zamri Abdul Talib, Mohd Afendy Lau, Han Yih Shamsudin, Mariana Nor Abdul Rahim, Raha |
| author_facet | Mohd Saad, Suria Abdul Kadir, Adlin Azlina Ishak, Zamri Abdul Talib, Mohd Afendy Lau, Han Yih Shamsudin, Mariana Nor Abdul Rahim, Raha |
| author_sort | Mohd Saad, Suria |
| collection | UPM |
| description | The SYBR Green I real-time PCR assay was used to quantify E. coli O157:H7 in various meat samples. Primers were designed to amplify and quantify the structural flagella (fliC) gene of E. coli O157:H7 in a single reaction. The primer specificity was confirmed with DNA from an ATCC culture of E. coli O157:H7 EDL933 as positive control, autoclaved E. coli O157:H7 EDL933 as negative control (NC) and nuclease free water as non template control (NTC). A direct correlation was determined between the fluorescence threshold (Ct) and the starting quantity of E. coli O157:H7 DNA. A detection limit of 4.71 x 10–2 ng/ μl of E. coli O157:H7 DNA equivalent to approximately 1.4 x 10–2 CFU of E. coli O157:H7 ml–1 based on plate counts was determined. Quantification of E. coli O157:H7 in Australian and Malaysian beef, chicken meat, burger and minced beef from the markets was possible when DNA quantity was as low as 1.0 x 10–2 ng/μl. These results indicated that the developed PCR assay was suitable for quantitative determination of E. coli O157:H7 in meat samples. |
| first_indexed | 2024-03-06T09:06:36Z |
| format | Article |
| id | upm.eprints-49301 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T09:06:36Z |
| publishDate | 2012 |
| publisher | Malaysian Agricultural Research and Development Institute (MARDI) |
| record_format | dspace |
| spelling | upm.eprints-493012016-12-30T02:30:53Z http://psasir.upm.edu.my/id/eprint/49301/ Real-time PCR for detection of fliC gene of E. coli O157:H7 in beef and chicken meat Mohd Saad, Suria Abdul Kadir, Adlin Azlina Ishak, Zamri Abdul Talib, Mohd Afendy Lau, Han Yih Shamsudin, Mariana Nor Abdul Rahim, Raha The SYBR Green I real-time PCR assay was used to quantify E. coli O157:H7 in various meat samples. Primers were designed to amplify and quantify the structural flagella (fliC) gene of E. coli O157:H7 in a single reaction. The primer specificity was confirmed with DNA from an ATCC culture of E. coli O157:H7 EDL933 as positive control, autoclaved E. coli O157:H7 EDL933 as negative control (NC) and nuclease free water as non template control (NTC). A direct correlation was determined between the fluorescence threshold (Ct) and the starting quantity of E. coli O157:H7 DNA. A detection limit of 4.71 x 10–2 ng/ μl of E. coli O157:H7 DNA equivalent to approximately 1.4 x 10–2 CFU of E. coli O157:H7 ml–1 based on plate counts was determined. Quantification of E. coli O157:H7 in Australian and Malaysian beef, chicken meat, burger and minced beef from the markets was possible when DNA quantity was as low as 1.0 x 10–2 ng/μl. These results indicated that the developed PCR assay was suitable for quantitative determination of E. coli O157:H7 in meat samples. Malaysian Agricultural Research and Development Institute (MARDI) 2012 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/49301/1/Real-time%20PCR%20for%20detection%20of%20fliC%20gene%20of%20E.%20coli%20O157H7%20in%20beef%20and%20chicken%20meat.pdf Mohd Saad, Suria and Abdul Kadir, Adlin Azlina and Ishak, Zamri and Abdul Talib, Mohd Afendy and Lau, Han Yih and Shamsudin, Mariana Nor and Abdul Rahim, Raha (2012) Real-time PCR for detection of fliC gene of E. coli O157:H7 in beef and chicken meat. Journal of Tropical Agriculture and Food Science, 40 (1). pp. 81-88. ISSN 1394-9829; ESSN: 2289-9650 http://ejtafs.mardi.gov.my/index.php/publication/issues/archive/64-2012/volume-40-no1/249-400109 |
| spellingShingle | Mohd Saad, Suria Abdul Kadir, Adlin Azlina Ishak, Zamri Abdul Talib, Mohd Afendy Lau, Han Yih Shamsudin, Mariana Nor Abdul Rahim, Raha Real-time PCR for detection of fliC gene of E. coli O157:H7 in beef and chicken meat |
| title | Real-time PCR for detection of fliC gene of E. coli O157:H7 in beef and chicken meat |
| title_full | Real-time PCR for detection of fliC gene of E. coli O157:H7 in beef and chicken meat |
| title_fullStr | Real-time PCR for detection of fliC gene of E. coli O157:H7 in beef and chicken meat |
| title_full_unstemmed | Real-time PCR for detection of fliC gene of E. coli O157:H7 in beef and chicken meat |
| title_short | Real-time PCR for detection of fliC gene of E. coli O157:H7 in beef and chicken meat |
| title_sort | real time pcr for detection of flic gene of e coli o157 h7 in beef and chicken meat |
| url | http://psasir.upm.edu.my/id/eprint/49301/1/Real-time%20PCR%20for%20detection%20of%20fliC%20gene%20of%20E.%20coli%20O157H7%20in%20beef%20and%20chicken%20meat.pdf |
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