Loop-Mediated Isothermal Amplification (LAMP) for the Detection of Horse Meat in Meat and Processed Meat Products
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Loop-Mediated Isothermal Amplification (LAMP) for the Detection of Horse Meat in Meat and Processed Meat Products Celine Zahradnik & Roland Martzy & Robert L. Mach & Rudolf Krska & Andreas H. Farnleitner & Kurt Brunner
Received: 4 September 2014 / Accepted: 15 December 2014 # Springer Science+Business Media New York 2014
Abstract In this study, a simple and rapid high-throughput method for the detection of horse meat in processed food products is described. Specific loop-mediated isothermal amplification (LAMP) primers were designed to target the mitochondrial genome of horse (Equus caballus). No crossreactions were observed for beef, pork, and chicken. Sensitivity tests showed reliable detection of 0.1 ng of extracted horse DNA. Spiking experiments were performed to show that the assay is capable of detecting 0.1 % horse meat in prepared model sausages, independent from their cooking time. Additionally, five different commercial horse meat products were analyzed to ensure the robustness of the assay when applied to varying food matrices. All experiments were performed on a heating block followed by visual detection using an intercalating dye. Results were confirmed by real-time fluorescence monitoring using a thermal cycler and compared to a previously published real-time PCR assay. In conclusion, this method is a good candidate for the simple and efficient testing of horse meat in food-products in the future.
Electronic supplementary material The online version of this article (doi:10.1007/s12161-014-0072-8) contains supplementary material, which is available to authorized users. C. Zahradnik : R. Martzy : K. Brunner (*) Institute of Chemical Engineering, IFA-Tulln, Center for Analytical Chemistry, Vienna University of Technology, Konrad Lorenz Str. 20, 3430 Tulln, Austria e-mail: [email protected] R. L. Mach : A. H. Farnleitner Institute of Chemical Engineering, Gene Technology Group, Vienna University of Technology, Gumpendorfer Str. 1a, 1060 Vienna, Austria R. Krska Department IFA-Tulln, Center for Analytical Chemistry, University of Natural Resources and Applied Life Sciences Vienna, Konrad Lorenz Str. 20, 3430 Tulln, Austria
Keywords Loop-mediated isothermal amplification . Horse meat . Visual detection . Sausages . Meat species identification
Introduction In 2012 and 2013, the European Commission performed official controls in several European Union member states, revealing the addition of horse meat in pre-packaged food products labelled as 100 % beef. Followed by these discoveries, the meat adulteration scandal was sparked in January 2013, especially in the UK, Ireland, and Germany, where most adulterated products were found (European Commission 2013). Although horse meat is considered a delicacy in some countries, the European Commission raised concerns about these findings. Since labelling requirements were violated, the fraudulent nature of this matter can be assumed. Apart from that, the consumption of non-declared horse meat might also pose a health risk to consumers. Phenylbutazone is a
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