A Method for Identification of Adulteration in Meat Products by DNA Extraction-Free qPCR Lyophilized Reagents

Abstract:

[Objective] This study aims to achieve rapid quantitative detection of beef- and duck-derived ingredients in meat products. [Method] The DNA extraction-free method was combined with fluorescent quantitative PCR (qPCR) lyophilized reagents was developed. The DNA extraction-free method was employed to prepare the template for qPCR. The premixed qPCR reagents were prepared by lyophilization. A portable qPCR system was used for amplification with the lyophilized reagents. [Result] The DNA extraction-free method with simplified sample extraction and purification steps achieved the detection within 10 min. The method combining the DNA extraction-free DNA method and qPCR lyophilized reagents can produce results within 45 min, with the limit of detection being 1 pg/μL. The premixed qPCR lyophilized reagents can be stored at 4 ℃ for 6 months, at room temperature for 4 months, and at 37 ℃ for 14 d. [Conclusion] The qPCR lyophilized reagents are appropriate for a portable qPCR system and can thus achieve rapid identification of adulteration in meat products.