DETECTION OF PIG MEAT, LIVER AND LARD IN BEEF BY CE-SSCP

Identification of animal species from foodstuffs is important in order to identify frauds to prevent substitutions and admixtures in animal products. In this paper we demonstrate the identification of cattle and pig species by polymerase chain reaction (PCR) capillary electrophoresis - single stranded conformation polymorphism (CE-SSCP) method. The procedure is based on the amplification of the 12S rRNA gene encoded in the mitochondrial DNA (mtDNA). Since mtDNA copy number is highly tissue dependent mixtures of different pig tissues in cattle meat were prepared, at a concentration of 1, 5, 10, 20 w/w% of pig lard, liver and loin. It was determined that regardless the tissue type pig DNA can be detected by CE-SSCP at each contamination level

Detection of DNA mutations by PCR-TTGE method

In our study PCR-temporal temperature gelelectrophoresis (TTGE) and MeltINGENY bioinformatic program were used to analyse the mutations in the genes of melanocortin-1 receptor (MC1R) and pituitary adenylate-cyclase activating polypeptide (PACAP) in cattle. Amplification of target DNA by PCR was performed with GC-clamp primers and non-GC-clamp primers in simplex PCR reactions. The fragments were separated by denaturing polyacrylamide gelelectrophoresis (denaturing agents: high temperature, urea) after PCR reactions.MC1R homozygous individuals were used for the reaction. We concluded that MeltINGENY program makes the decision and detection system easier, and more simple as the melting profile of target sequence is determined by the software. In case of MC1R gene, PCR-TTGE method is appropriate for SNP detection, however PACAP gene polymorphism can not be identified by the method, because PACAP mutations are not included in melting domains, therefore PCR-TTGE cannot detect them

Search for polymorphism in exon 5 of cattle pituitary adenylate cyclase activating polypeptide gene

Pituitary adenylate cyclase-activating polypeptide is a neuropeptide expressed primarily in the hypothalamus and in other tissues, which divergent and extensive physiological functions are proved. Large number of SNPs in PACAP are published involved SNP that is associated with phenotypic trait of cattle as well. Hungarian Grey, Hungarian Simmental, Holstein, Charolais and Angus cattle breeds were involved in this study to search for polymorphism in exon 5 1–391 bp and G/A transition. Our results of PCR RFLP and PCR SSCP did not prove the occurrence neither G/A transition, nor any other SNP in cattle breeds involved