Development of a direct loop-mediated isothermal amplification (LAMP) assay for rapid and simple on-site detection of chicken in processed meat products

Abstract

We developed a direct loop-mediated isothermal amplification (LAMP) assay targeting the mitochondrial 16S rRNA gene to detect chicken in processed meat products. This rapid on-site detection method was developed using a portable fluorescence device and the LAMP assay with direct amplification. Chicken-specific LAMP primer set was designed and its specificity confirmed against 24 other animal species. The limit of detection (LOD) of the chicken-specific LAMP assay for chicken DNA was 10 fg. To evaluate the sensitivity of the direct LAMP assay, different percentages of chicken meat prepared with pork or beef were prepared. Heat- and pressure-treated meat mixtures were used to assess the applicability of this assay. LODs of the direct LAMP assay for the chicken/pork and chicken/beef mixtures were 0.1% for raw meat and 1% for heat- and pressure-treated meat. The direct LAMP assay was applied to 19 commercially available processed meat products. This assay successfully amplified the target sequence and identified the presence of chicken meat in processed meat products within approximately 30 min. This direct LAMP assay is a rapid and simple method for on-site detection of chicken meat in commercial products to monitor the authentication of chicken in meat products.