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Real-time PCR for detection of fliC gene of E. coli O157:H7 in beef and chicken meat


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


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.

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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
Faculty of Medicine and Health Science
Publisher: Malaysian Agricultural Research and Development Institute (MARDI)
Keywords: Real-time PCR; E. coli O157:H7; Meat; FliC gene
Depositing User: Nabilah Mustapa
Date Deposited: 30 Dec 2016 02:30
Last Modified: 30 Dec 2016 02:30
URI: http://psasir.upm.edu.my/id/eprint/49301
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