Species Authentication of Common Meat Based on PCR Analysis of the Mitochondrial COI Gene

Adulteration of meat products and costly animal-derived commodities with their inferior/cheaper counterparts is a grievous global problem. Species authentication is still technical challenging, especially to those deep processed products. The present study described the design of seven sets of species-specific primer based on a high heterozygous region of mitochondrial cytochrome c oxidase subunit I (COI) gene. These primers were proven to have high species specificity and no cross-reactions and unexpected products to different DNA source. Multiplex PCR assay was achieved for rapid and economical identification of four commonly consumed meats (pork, beef, chicken, and mutton). The conventional PCR assay was sensitive down to 0.001 ng of DNA template in the reactant. The developed method was also powerful in detecting as low as 0.1-mg adulterated pork (0.05 % in wt/wt) in an artificial counterfeited mutton. Validation test showed that the assay is specific, reproducible, and robust in commercial deep processed meats, leatherware, and feather commodities. This proposed method will be greatly beneficial to the consumers, food industry, leather, and feather commodity manufacture.

[1]  K. Arman,et al.  A highly sensitive and specific tetraplex PCR assay for soybean, poultry, horse and pork species identification in sausages: development and validation. , 2014, Meat science.

[2]  Phuvadol Thanakiatkrai,et al.  Direct-multiplex PCR assay for meat species identification in food products. , 2014, Food chemistry.

[3]  D. Swayne Microassay for measuring thermal inactivation of H5N1 high pathogenicity avian influenza virus in naturally infected chicken meat. , 2006, International journal of food microbiology.

[4]  K. Uddin,et al.  Species Authentication Methods in Foods and Feeds: the Present, Past, and Future of Halal Forensics , 2012, Food Analytical Methods.

[5]  B. Al-Safadi,et al.  Identification of meat species by PCR-RFLP of the mitochondrial COI gene. , 2012, Meat science.

[6]  Masato Saito,et al.  Meat species identification based on the loop mediated isothermal amplification and electrochemical DNA sensor , 2010 .

[7]  Z. Kesmen,et al.  PCR assay for the identification of animal species in cooked sausages. , 2007, Meat science.

[8]  Wentao Xu,et al.  A novel common primer multiplex PCR (CP-M-PCR) method for the simultaneous detection of meat species , 2009 .

[9]  A. Raha,et al.  Analysis of raw meats and fats of pigs using polymerase chain reaction for Halal authentication. , 2005, Meat science.

[10]  A. Gulluce,et al.  Identification of meat species by TaqMan-based real-time PCR assay. , 2009, Meat science.

[11]  Hua H. Wang,et al.  Real-time Polymerase Chain Reaction for the Food Microbiologist: Technologies, Applications, and Limitations , 2006 .

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

[13]  M. G. Garner,et al.  How do resources influence control measures during a simulated outbreak of foot and mouth disease in Australia? , 2014, Preventive veterinary medicine.

[14]  Wan Ji,et al.  A method for quantifying mixed goat cashmere and sheep wool. , 2011, Forensic science international.

[15]  P S Girish,et al.  Sequence analysis of mitochondrial 12S rRNA gene can identify meat species. , 2004, Meat science.

[16]  E. Joseph,et al.  Characterization of 12S rRNA gene for meat identification of common wild and domestic small herbivores as an aid to wildlife forensic. , 2013 .

[17]  Chi Zhang Semi-nested multiplex PCR enhanced method sensitivity of species detection in further-processed meats , 2013 .

[18]  Daya Luo,et al.  Detection of single nucleotide polymorphisms by PCR conformation-difference gel electrophoresis , 2012, Biotechnology Letters.

[19]  A. Moussavi,et al.  Fraud identification in industrial meat products by multiplex PCR assay. , 2009 .

[20]  V. S. Melo,et al.  Authentication of a traditional game meat sausage (Alheira) by species-specific PCR assays to detect hare, rabbit, red deer, pork and cow meats , 2014 .

[21]  Isabel Mafra,et al.  A SYBR Green real-time PCR assay to detect and quantify pork meat in processed poultry meat products. , 2013, Meat science.

[22]  P. Hernández,et al.  Technical note: detection of chicken, turkey, duck, and goose tissues in feedstuffs using species-specific polymerase chain reaction. , 2007, Journal of animal science.

[23]  T. Tsutsui,et al.  A quantitative assessment of the risk of exposure to bovine spongiform encephalopathy via meat-and-bone meal in Japan. , 2006, Preventive veterinary medicine.

[24]  N. Mucci,et al.  Authentication of meat from game and domestic species by SNaPshot minisequencing analysis. , 2008, Meat science.

[25]  J. Chisholm,et al.  Effect of heat and pressure processing on DNA fragmentation and implications for the detection of meat using a real-time polymerase chain reaction , 2006, Food additives and contaminants.

[26]  Yong-Gang Yao,et al.  Species authentication of commercial beef jerky based on PCR-RFLP analysis of the mitochondrial 12S rRNA gene. , 2010, Journal of genetics and genomics = Yi chuan xue bao.

[27]  C. Stamatis,et al.  A new set of 16S rRNA universal primers for identification of animal species , 2014 .