Identification for meat adulteration (pork, beef, sheep and duck) in foodstuff by microfluidic chip-based real-time fluorescent LAMP

Meat adulteration has become one of the most concerning issues in the global food industry, affecting economic investment, public health and so on. In this study, a real-time fluorescent loop-mediated isothermal amplification (LAMP)-microfluidic assay was developed to simultaneously detect pork, beef, sheep and duck in foodstuffs using species-specific primers targeting the mitochondrial cytochrome b gene and cytochrome c oxidase subunit gene. Various primers were precoated in different reaction chambers of the real-time fluorescent LAMP-microfluidic chip, and the entire amplification was completed within 30 min. The real-time fluorescent LAMP-microfluidic assay exhibited good specificity and high sensitivity, with the limit of detection reaching 0.1% (w/w) in simu-lated adulterated samples, satisfying authentication requirements. The accuracy of this novel method was validated using 33 commercial samples and was comparable with that of the typical real-time qPCR assay. In summary, the real-time fluorescent LAMP-microfluidic assay can simultaneously detect pork, beef, sheep and duck in commercial products to verify the label certification of meat and prevent food adulteration.