Species specific polymerase chain reaction (PCR) assay for identification of pig (Sus domesticus) meat

A highly specific single step polymerase chain reaction (PCR) is described for the detection of pig ( Sus  domesticus ) meat. A PCR assay was successfully optimized for amplification of 629 and 322-bp DNA fragment extracted from pig meat using designed species-specific primer pairs based on mitochondrial D-loop and 12S ribosomal ribonucleic acid (rRNA) gene, respectively. The optimized PCR assay was subsequently validated for its specificity with deoxyribonucleic acid (DNA) extracted from cattle, buffalo, sheep, goat and pig. PCR amplification of target DNA with pig-specific primers was repeated 15 times, with consistent results  observed. The specificity of pig-specific PCR provides a valuable tool for identification of pig meat and to avoid its fraudulent substitution and adulteration. Key words: Pig meat, adulteration, polymerase chain reaction (PCR), mitochondrial D-Loop, 12S ribosomal ribonucleic acid (rRNA) gene.

[1]  M Chévremont,et al.  Mitochondrial DNA , 2009, Encyclopedia of Biometrics.

[2]  D. Sharma,et al.  Differentiation of meat species by means of Polymerase Chain Reaction technique , 2008 .

[3]  N. Karabasanavar Molecular identification of animal species using Polymerase Chain Reaction based techniques , 2008 .

[4]  Paolo Ajmone-Marsan,et al.  Standard and Light-Cycler PCR methods for animal DNA species detection in animal feedstuffs , 2008 .

[5]  A. Arslan,et al.  Identification of Meat Species by Polymerase Chain Reaction (PCR) Technique , 2007 .

[6]  T. García,et al.  Mitochondrial markers for the detection of four duck species and the specific identification of Muscovy duck in meat mixtures using the polymerase chain reaction. , 2007, Meat science.

[7]  M. Saini,et al.  Characterisation of peacock (Pavo cristatus) mitochondrial 12S rRNA sequence and its use in differentiation from closely related poultry species , 2007, British poultry science.

[8]  P. V. Ravindra,et al.  Precise and rapid identification of ovine-caprine origin of meat by specific PCR assay. , 2007 .

[9]  S. Wasser,et al.  The potential of mitochondrial DNA markers and polymerase chain reaction-restriction fragment length polymorphism for domestic and wild species identification , 2006 .

[10]  Dietrich Maede A strategy for molecular species detection in meat and meat products by PCR-RFLP and DNA sequencing using mitochondrial and chromosomal genetic sequences , 2006 .

[11]  A. Arslan,et al.  Effect of method of cooking on identification of heat processed beef using polymerase chain reaction (PCR) technique. , 2006, Meat science.

[12]  Belinda Vallejo-Cordoba,et al.  Capillary electrophoresis for the analysis of meat authenticity. , 2005, Journal of separation science.

[13]  P S Girish,et al.  Meat species identification by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene. , 2005, Meat science.

[14]  Allan C. Wilson,et al.  Mitochondrial DNA sequences of primates: Tempo and mode of evolution , 2005, Journal of Molecular Evolution.

[15]  B. Brenig,et al.  Diagnostic polymorphisms in the mitochondrial cytochrome b gene allow discrimination between cattle, sheep, goat, roe buck and deer by PCR-RFLP , 2004, BMC Genetics.

[16]  I. Giovannacci,et al.  Species identification of meat products by ELISA , 2004 .

[17]  T. García,et al.  PCR identification of beef, sheep, goat, and pork in raw and heat-treated meat mixtures. , 2004, Journal of food protection.

[18]  M. Hajmeer,et al.  Spinal cord tissue detection in comminuted beef: comparison of two immunological methods. , 2003, Meat science.

[19]  Chich-Sheng Lin,et al.  Establishment and application of a fluorescent polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identifying porcine, caprine, and bovine meats. , 2003, Journal of agricultural and food chemistry.

[20]  H. Rehbein,et al.  On the specificity of tuna-directed primers in PCR-SSCP analysis of fish and meat , 2001 .

[21]  R. Osta,et al.  Technical note: A quick and more sensitive method to identify pork in processed and unprocessed food by PCR amplification of a new specific DNA fragment. , 2001, Journal of animal science.

[22]  S. Comincini,et al.  Identification of species in animal feedstuffs by polymerase chain reaction--restriction fragment length polymorphism analysis of mitochondrial DNA. , 2001, Journal of agricultural and food chemistry.

[23]  J. Lenstra,et al.  On the origin of meat - DNA techniques for species identification in meat products , 2001 .

[24]  P. Zanelli,et al.  Identification of anchovy (Engraulis encrasicholus L.) and gilt sardine (Sardinella aurita) by polymerase chain reaction, sequence of their mitochondrial cytochrome b gene, and restriction analysis of polymerase chain reaction products in semipreserves. , 2001, Journal of agricultural and food chemistry.

[25]  J. Murby,et al.  Evaluation of a DNA fingerprinting method for determining the species origin of meats. , 2000, Meat science.

[26]  F C Chen,et al.  Detection of pork in heat-processed meat products by monoclonal antibody-based ELISA. , 2000, Journal of AOAC International.

[27]  J. Lenstra,et al.  On the origin of meat , 1999 .

[28]  J. Rentsch,et al.  PCR-RFLP analysis of mitochondrial DNA: a reliable method for species identification. , 1999, Journal of agricultural and food chemistry.

[29]  S Muroya,et al.  A quick and simple method for the identification of meat species and meat products by PCR assay. , 1999, Meat science.

[30]  S. Pääbo,et al.  Nuclear insertion sequences of mitochondrial DNA predominate in hair but not in blood of elephants , 1999, Molecular ecology.

[31]  T. Matsunaga,et al.  Effects of Processing Conditions on Species Identification of Meat Products. , 1999 .

[32]  J. Lenstra,et al.  Species identification in meat by using PCR-generated satellite probes , 1998, Journal of Industrial Microbiology and Biotechnology.

[33]  R. Ball,et al.  Methodology and development of prediction equations for the determination of pork substitution in veal , 1997 .

[34]  P. Brandt,et al.  Identification of the species origin of highly processed meat products by mitochondrial DNA sequences. , 1995, PCR methods and applications.

[35]  W. Davidson,et al.  FINS (forensically informative nucleotide sequencing): a procedure for identifying the animal origin of biological specimens. , 1992, BioTechniques.

[36]  P. G. Stiles,et al.  Liquid chromatographic identification of meats. , 1988, Journal - Association of Official Analytical Chemists.

[37]  C. Baur,et al.  Spezifizierung hitzedenaturierter Fleischproben durch DNA-Analyse , 1987 .