Identification of Species of Meat Origin using ATPase Gene Variability by the Polymerase Chain Reaction

Abstract

Food authenticity is currently an issue of major concern for food authorities, since incorrect labelling of animal foods may have remarkable negative consequences. To circumvent this problem, DNA based method had been utilised. The present study was carried out for the detection of meat species by using variability in ATPase Subunit 6 and 8 genes by multiplex PCR. Meat samples from cattle, buffalo, sheep, goat, chicken and dog were utilized for molecular analysis. KAPA express extract kit was used to extract DNA from meat samples. Sequences among mitochondrial ATPase Subunit 6 and 8 genes were targeted for species-specific amplifications. The specificity of the primers was checked by using the Basic Local Alignment Search Tool (BLAST) software. The designed primers yielded specific amplification of 65, 107, 144, 186, 200 and 232 bp for Goat, Sheep, Buffalo, Dog, Poultry and Cattle respectively. Further to detect the sensitivity of the assay different level of meat mixture of 16, 8, 4, 2, 1 and 0.1% was formulated by adding pork. The assay successfully identified the presence of meat at the level of 1%. In the current assay, cooked and putrefied meat samples also showed successful amplification of the DNA, so this assay is useful in the detection of meat adulteration.