Citation
Zamri Ishak, . and Adlin Azlina Abdul Kadir, . and Lau Han Yih, . and Mariana Nor Shamsudin, . and Suria Mohd Saad, . and Raha Abd Rahim, . and Mohd Afendy Abdul Talib, . Real-time PCR for detection of fliC gene of E. coli O157:H7 in beefand chicken meat. pp. 81-88. ISSN 1394-9829
Abstract
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/l 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/l. These results indicated that the developed PCR assay was suitable for quantitative determination of E. coli O157:H7 in meat samples.
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Abstract
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/l 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/l. These results indicated that the developed PCR assay was suitable for quantitative determination of E. coli O157:H7 in meat samples.
Additional Metadata
| Item Type: | Article |
|---|---|
| AGROVOC Term: | Chicken meat |
| AGROVOC Term: | PCR |
| AGROVOC Term: | Genes |
| AGROVOC Term: | Escherichia coli |
| AGROVOC Term: | Chicken meat |
| AGROVOC Term: | Beef |
| AGROVOC Term: | Polymerase chain reaction |
| AGROVOC Term: | DNA |
| AGROVOC Term: | Nucleases |
| AGROVOC Term: | Foodborne diseases |
| Depositing User: | Ms. Suzila Mohamad Kasim |
| Last Modified: | 24 Apr 2025 06:29 |
| URI: | http://webagris.upm.edu.my/id/eprint/24488 |
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