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Detection of horse DNA by loop-mediated isothermal amplification (LAMP) seems one of the most promising methods to meet the criteria of fast, robust, cost efficient, specific, and sensitive on-site detection. In the present study an assessment of the specificity and sensitivity of the LAMP horse screening assay was made and outcomes were compared with the EURL-AP (European Union Reference laboratory for Animal Proteins in feeding stuffs) qPCR method. The specificity was tested with DNA samples from seven other species. The sensitivity of the LAMP assay was subsequently challenged with different percentages of horse DNA in cattle DNA and different percentages of horse meat in cattle meat. Both qPCR and LAMP were able to reliably detect horse DNA or meat below 0.1%, but LAMP was able to do so in less than 30 min. The DNA of other species did not result in a response in the LAMP horse assay. These features show that the LAMP method is fast, specific, and sensitive. Next, 69 processed meat samples were screened for the presence of horse DNA. The results showed that the LAMP horse assay, combined with a simple and fast on-site DNA extraction method, results in similar outcomes as the EURL-AP qPCR method and is thus a promising screening assay to be used outside the laboratory. Only samples that are screened on-site as suspect in the LAMP horse assay, need to be brought to the laboratory for confirmation with the more laborious EURL-AP qPCR reference method.
- Horse meat
- Loop-mediated isothermal amplification (LAMP)
- On-site detection
- Rapid method