title: Authentication of beef, carabeef, chevon, mutton and pork by a PCR-RFLP assay of mitochondrial cytb gene.
abstract: Authentication of meat assumes significance in view of religious, quality assurance, food safety, public health, conservation and legal concerns. Here, we describe a PCR-RFLP (Polymerase Chain Reaction- Restriction Fragment Length Polymorphism) assay targeting mitochondrial cytochrome-b gene for the identification of meats of five most common food animals namely cattle, buffalo, goat, sheep and pig. A pair of forward and reverse primers (VPH-F & VPH-R) amplifying a conserved region (168-776 bp) of mitochondrial cytochrome-b (cytb) gene for targeted species was designed which yielded a 609 bp PCR amplicon. Further, restriction enzyme digestion of the amplicons with Alu1 and Taq1 restriction enzymes resulted in a distinctive digestion pattern that was able to discriminate each species. The repeatability of the PCR-RFLP assay was validated ten times with consistent results observed. The developed assay can be used in routine diagnostic laboratories to differentiate the meats of closely related domestic livestock species namely cattle from buffalo and sheep from goat.
authors.affiliation: Department of Veterinary Public Health, College of Veterinary & Animal Sciences, G.B. Pant University of Agriculture & Technology, Pantnagar, 263 145 Uttarakhand India ; Department of Veterinary Public Health & Epidemiology, College of Veterinary Sciences & Animal Husbandry, R. K. Nagar, Agartala, 799008 Tripura India.
authors.forename: Deepak
authors.initials: D
authors.lastname: Kumar
authors.affiliation: Department of Veterinary Public Health, College of Veterinary & Animal Sciences, G.B. Pant University of Agriculture & Technology, Pantnagar, 263 145 Uttarakhand India.
authors.forename: S P
authors.initials: SP
authors.lastname: Singh
authors.affiliation: Department of Veterinary Public Health, Bombay Veterinary College, Mumbai, 400012 Maharashtra India.
authors.forename: Nagappa S
authors.initials: NS
authors.lastname: Karabasanavar
authors.affiliation: Department of Veterinary Microbiology & Immunology, College of Veterinary & Animal Sciences, G.B. Pant University of Agriculture & Technology, Pantnagar, 263 145 Uttarakhand India.
authors.forename: Rashmi
authors.initials: R
authors.lastname: Singh
authors.affiliation: Indian Veterinary Research Institute, Regional Station, Palampur, 176061 Himachal Pradesh India.
authors.forename: V
authors.initials: V
authors.lastname: Umapathi
keywords: Adulteration; AluI; Cytochrome-b gene; Meat; PCR-RFLP; TaqI