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Loop-Mediated Isothermal Amplification (LAMP) for the Detection of Horse Meat in Meat and Processed Meat Products

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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 cross-reactions 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.

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Acknowledgments

This work was financially supported by the Federal Country Lower Austria in cooperation with the European Regional Development Fund (ERDF).

Conflict of interest

Celine Zahradnik declares that she has no conflict of interest. Roland Martzy declares that he has no conflict of interest. Robert L. Mach declares that he has no conflict of interest. Rudolf Krska declares that he has no conflict of interest. Andreas H. Farnleitner declares that he has no conflict of interest. Kurt Brunner declares that he has no conflict of interest.

Compliance with Ethics Requirements

This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. Institute of Chemical Engineering, IFA-Tulln, Center for Analytical Chemistry, Vienna University of Technology, Konrad Lorenz Str. 20, 3430, Tulln, Austria

    Celine Zahradnik, Roland Martzy & Kurt Brunner

  2. Institute of Chemical Engineering, Gene Technology Group, Vienna University of Technology, Gumpendorfer Str. 1a, 1060, Vienna, Austria

    Robert L. Mach & Andreas H. Farnleitner

  3. Department IFA-Tulln, Center for Analytical Chemistry, University of Natural Resources and Applied Life Sciences Vienna, Konrad Lorenz Str. 20, 3430, Tulln, Austria

    Rudolf Krska

Authors
  1. Celine Zahradnik
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  2. Roland Martzy
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  3. Robert L. Mach
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  4. Rudolf Krska
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  5. Andreas H. Farnleitner
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  6. Kurt Brunner
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Corresponding author

Correspondence to Kurt Brunner.

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Table S1

The presence of amplifyable DNA was proven via real-time PCR using a universal primer set for vertebrates targeting parts of the 12S rRNA, as previously published (Kitano et al. 2007), n = 10. (DOC 11 kb)

Figure S1

Agarose gel electrophoresis of the amplification products, resulting in a pattern typical for the various-sized LAMP products. Lane 1-10 - horse meat [10 ng/μL], lanes 11-15 - beef [10 ng/μL], lanes 16-20 - non-template control (NTC). (DOC 255 kb)

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Zahradnik, C., Martzy, R., Mach, R.L. et al. Loop-Mediated Isothermal Amplification (LAMP) for the Detection of Horse Meat in Meat and Processed Meat Products. Food Anal. Methods 8, 1576–1581 (2015). https://doi.org/10.1007/s12161-014-0072-8

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  • DOI: https://doi.org/10.1007/s12161-014-0072-8

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