Rapid detection of porcine DNA in processed food samples using a streamlined DNA extraction method combined with the SYBR Green real-time PCR assay. (30th March 2020)

• Record Type: Journal Article
• Authors: Tan, Lee Lee; Ahmed, Siti Aminah; Ng, Siew Kit; Citartan, Marimuthu; Raabe, Carsten A.; Rozhdestvensky, Timofey S.; Tang, Thean Hock
• Is Part Of: Food chemistry. Volume 309(2020)
• Publication Date: 2020-03-30
• Subjects: Genomic DNA extraction -- Food authentication -- Sus scrofa -- Adulteration with pork -- SYBR Green PCR -- Melting curve analysis; Food -- Analysis -- Periodicals; Food -- Composition -- Periodicals; 664
• DOI: 10.1016/j.foodchem.2019.125654
• Languages: English
• ISSNs: 0308-8146

Abstract

Highlights: Quantitative PCR assay to detect food adulterated with pork was established. Amplification of porcine-specific LINE-1 and internal control in a single tube was achieved. Rapid and efficient DNA extraction from samples and PCR analysis were completed in 10 min. Detection threshold of 0.001% pork within meat mixtures was achieved. Robustness was demonstrated with analysis of 121 commercial meat-based food products. Abstract: A specialized DNA extraction method and a SYBR Green quantitative polymerase chain reaction (SyG-qPCR) assay were combined to generate a ready-to-use kit for rapid detection of porcine admixtures in processed meat products. Our qPCR assay utilized repetitive LINE-1 elements specific to the genome of Sus scrofa domesticus (pig) as a target and incorporated internal controls. We improved the genomic DNA extraction method, and reduced extraction times to the minimum. The method was validated for specificity, sensitivity (0.001% w/w) and robustness, and values were compared with those of a commercially available kit. We also tested our method using 121 processed food products and consistently detected amplification only in samples containing pork. Due to its efficiency and cost-effectiveness, our method represents a valuable new method for detecting food adulteration with pork that is superior to existing quality control approaches.