Detection and quantification of beef and pork materials in meat products by duplex droplet digital PCR

Meat products often consist of meat from multiple animal species, and inaccurate food product adulteration and mislabeling can negatively affect consumers. Therefore, a cost-effective and reliable method for identification and quantification of animal species in meat products is required. In this study, we developed a duplex droplet digital PCR (dddPCR) detection and quantification system to simultaneously identify and quantify the source of meat in samples containing a mixture of beef (Bos taurus) and pork (Sus scrofa) in a single digital PCR reaction tube. Mixed meat samples of known composition were used to test the accuracy and applicability of this method. The limit of detection (LOD) and the limit of quantification (LOQ) of this detection and quantification system were also identified. We conclude that our dddPCR detection and quantification system is suitable for quality control and routine analyses of meat products.

[1]  R. Köppel,et al.  Multiplex real-time PCR for the detection and quantification of DNA from beef, pork, chicken and turkey , 2008 .

[2]  Xin Fang,et al.  Detection of adulterated murine components in meat products by TaqMan© real-time PCR. , 2016, Food chemistry.

[3]  L. Fontanesi,et al.  A Next Generation Semiconductor Based Sequencing Approach for the Identification of Meat Species in DNA Mixtures , 2015, PloS one.

[4]  R. Köppel,et al.  Multiplex real-time PCR for the detection and quantification of DNA from beef, pork, horse and sheep , 2008 .

[5]  M. Shimokomaki,et al.  Hamburger meat identification by dot-ELISA. , 2000, Meat science.

[6]  J. Demarquoy Meat adulteration: the use of PCR. , 2013, Meat science.

[7]  J. Ofori,et al.  Detection of horse meat contamination in raw and heat-processed meat products. , 2014, Journal of agricultural and food chemistry.

[8]  B Brenig,et al.  Species identification and quantification in meat and meat products using droplet digital PCR (ddPCR). , 2015, Food chemistry.

[9]  Yicun Cai,et al.  Quantitative Analysis of Pork and Chicken Products by Droplet Digital PCR , 2014, BioMed research international.

[10]  Alison S. Devonshire,et al.  International Interlaboratory Digital PCR Study Demonstrating High Reproducibility for the Measurement of a Rare Sequence Variant. , 2017, Analytical chemistry.

[11]  R. Osta,et al.  Beef- and bovine-derived material identification in processed and unprocessed food and feed by PCR amplification. , 2002, Journal of agricultural and food chemistry.

[12]  Deepak Sharma,et al.  Identification of Species Origin of Meat and Meat Products on the DNA Basis: A Review , 2015, Critical reviews in food science and nutrition.

[13]  Benjamin J. Hindson,et al.  Evaluation of a Droplet Digital Polymerase Chain Reaction Format for DNA Copy Number Quantification , 2011, Analytical chemistry.

[14]  D. Tesfaye,et al.  Haplotype analysis of beta-actin gene for its association with sperm quality and boar fertility. , 2006, Journal of animal breeding and genetics = Zeitschrift fur Tierzuchtung und Zuchtungsbiologie.

[15]  F. Şahin,et al.  Detection of chicken and turkey meat in meat mixtures by using real-time PCR assays. , 2012, Journal of food science.

[16]  Rani Alex,et al.  Application of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cow Components Adulterated in Buffalo Milk/Meat , 2016, Molecular Biotechnology.

[17]  H. Barakat,et al.  Detection of pork adulteration in processed meat by species-specific PCR-QIAxcel procedure based on D-loop and cytb genes , 2014, Applied Microbiology and Biotechnology.

[18]  S. Singh,et al.  Detection of pork adulteration by highly-specific PCR assay of mitochondrial D-loop. , 2014, Food chemistry.

[19]  M. Cichna‐Markl,et al.  Development and validation of a TaqMan real-time PCR assay for the identification and quantification of roe deer (Capreolus capreolus) in food to detect food adulteration. , 2015, Food chemistry.

[20]  Jeff Mellen,et al.  High-Throughput Droplet Digital PCR System for Absolute Quantitation of DNA Copy Number , 2011, Analytical chemistry.

[21]  A. Parente,et al.  Myoglobin as marker in meat adulteration: a UPLC method for determining the presence of pork meat in raw beef burger. , 2013, Food chemistry.

[22]  S. Sheu,et al.  Development of a monoclonal antibody specific to cooked mammalian meats. , 1998, Journal of food protection.

[23]  F. Vogensen,et al.  Species determination - Can we detect and quantify meat adulteration? , 2009, Meat science.

[24]  H. Akiyama,et al.  A Real-Time Quantitative PCR Detection Method for Pork, Chicken, Beef, Mutton, and Horseflesh in Foods , 2007, Bioscience, biotechnology, and biochemistry.

[25]  Mojca Milavec,et al.  Quantitative Analysis of Food and Feed Samples with Droplet Digital PCR , 2013, PloS one.