Development and validation of a rapid kit for authenticity of murine meat in meat products with a species-specific PCR assay. Issue 4 (2nd April 2020)

• Record Type: Journal Article
• Authors: Duan, Siqi; Ai, Jin Xia; Sun, Liyuan; Gao, Lijun; Li, Mingcheng; Chen, Kun; Li, Dan
• Is Part Of: Food additives & contaminants. Volume 37:Issue 4(2020)
• Page Start: 552
• Page End: 560
• Publication Date: 2020-04-02
• Subjects: Authentication -- fraud -- murine-species ingredients -- common edible meat -- kit -- mitochondrial DNA; Food additives -- Periodicals; Food contamination -- Periodicals; 664.06
• DOI: 10.1080/19440049.2020.1718218
• Languages: English
• ISSNs: 1944-0049

Abstract

ABSTRACT: Adulteration of meat products with murine meat poses a huge threat to consumer health and leads to serious disruption in food markets. Species authentication of murine meat is still technically challenging. We, therefore, developed a species-specific PCR kit consisting of murine meat DNA extraction, PCR reaction and identifying systems. We designed novel universal primers targeting highly conserved region on mitochondrial cytochrome b gene ( cyt b) from four murines (lab rats, lab mice, wild rat and wild mice), as well as specific primers for meat from four widely consumed animal species, cattle, sheep, duck and donkey. Simultaneously, pasmid inserted specific cyt b fragment was cloned and used as the internal positve control in the kit. The kit parameters of specificity, sensitivity, stability and validity were determined using mimic counterfeiting meatball. The specificity of the DNA detection kit was 100% in authentication of the four fraudulent meats of cattle, sheep, duck and donkey mixed murine meat. The minimum detection limit of the sample DNA was 0.1 μg. The kit, which had freeze-thawed up to 20 times and stored for 1 year, also was powerful in detecting an amount of 0.1 mg in artificial counterfeited cattle, sheep, duck and donkey meat products. The murine-species DNA detection kit proposed in this study has proved to be a simple, accurate and effective assay, and can be applied to the identification of murine meat traces in common edible meat, to ensure the realisable implementation of meat product market supervision.