Visual loop-mediated isothermal amplification (LAMP) method for identification bovine and ovine gene in animal foodstuff

Abstract

Objective: The aim of this study was to develop an easy and highly sensitive visual loop-mediated isothermal amplification (LAMP) assay for specific detection of the bovine and ovine derived ingredient in foodstuff. Methodology: The primers designed based on different regions of mitochondrial DNA (CoI and Cyt b) for amplification of 184 and 206 bp fragments, respectively. The specificity of assay was cross tested with DNA of horse, pork and chicken, where amplification were observed in bovine and ovine without cross reactivity with others meat species. Results: The sensitivity was 5.0×10-3 ng for bovine, as same as the ovine genomic DNA. The detection limit of meat mixtures was 0.01% for bovine and 0.001% for ovine. Moreover, the LAMP assay was in concordance with the qPCR analysis in the test of practical samples. The visual LAMP assay can be considered as a valid tool for rapid detection of animal derived materials in meat products. Conclusion: It contributes to a future approach on food analysis by meeting the demands for quick and easy-to-perform analytical methods.