Rapid detection of porcine DNA in processed food samples using a streamlined DNA extraction method combined with the SYBR Green real-time PCR assay

Tan LL 1 , Ahmed SA 2 , Ng SK 3 , Citartan M 4 , Raabe CA 5 , Rozhdestvensky TS 6 Show all authors , Tang TH 7

Affiliations 

  • 1 Advanced Medical & Dental Institute, Universiti Sains Malaysia, Bertam, 13200 Kepala Batas, Malaysia. Electronic address: leelee.tan@aiesec.net
  • 2 Advanced Medical & Dental Institute, Universiti Sains Malaysia, Bertam, 13200 Kepala Batas, Malaysia. Electronic address: asiti2000@usm.my
  • 3 Advanced Medical & Dental Institute, Universiti Sains Malaysia, Bertam, 13200 Kepala Batas, Malaysia. Electronic address: skng@usm.my
  • 4 Advanced Medical & Dental Institute, Universiti Sains Malaysia, Bertam, 13200 Kepala Batas, Malaysia. Electronic address: citartan@usm.my
  • 5 Institute of Experimental Pathology, Centre for Molecular Biology of Inflammation (ZMBE), University of Muenster, Von-Esmarch-Strasse 56, D-48149 Muenster, Germany; Institute of Medical Biochemistry, Centre for Molecular Biology of Inflammation (ZMBE), University of Muenster, Von-Esmarch-Strasse 56, D-48149 Muenster, Germany; Brandenburg Medical School (MHB), Fehrbelliner Strasse 38, D-16816 Neuruppin, Germany. Electronic address: raabec@uni-muenster.de
  • 6 Core Facility Transgenic Animal and Genetic Engineering Models (TRAM), University Hospital of Muenster, D-48149 Muenster, Germany. Electronic address: rozhdest@uni-muenster.de
  • 7 Advanced Medical & Dental Institute, Universiti Sains Malaysia, Bertam, 13200 Kepala Batas, Malaysia. Electronic address: tangth@usm.my
Food Chem, 2020 Mar 30;309:125654.
PMID: 31678669 DOI: 10.1016/j.foodchem.2019.125654

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.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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