Using Mitochondrial Cytochrome B Gene for Authentication of Meats from Different Animal Sources

Abstract

The current study aimed to use the mitochondrial cytochrome b gene for authentication of meats from different animal sources, with special relevance to halal-related meat sources. In total, 35 raw meat samples were sampled, representing seven species, viz goats, sheep, pigs, cattle, camels, buffaloes and donkeys. Mitochondrial Cytochrome-b (mtCyt-b) gene sequences for the seven-investigated species were retrieved from GenBank database and aligned. Common universal forward oligonucleotide primer and specificreverse primer for each species were designed. DNA was extracted, PCR was performed using conventional assay and amplicons were resolved using standard gel electrophoresis. M-PCR proved to offer the following advantages: (i) PCR internal control: false negatives that pose potential problems in PCR are revealed in multiplex amplification since each amplicon allows internal control for the other amplified fragments. (ii) PCR efficiency of DNA template quality determination: with M-PCR technique degraded DNA templates show weaker signals for long bands than for short ones and for fall in amplification efficiency. (iii) PCR economic efficiency: by spotting multiple genes simultaneously, further information may be acquired from a single test run than would require more PCR reagents and preparation time. (iv) PCR rapidity: with M-PCR many meat products belonging to ruminants and non-ruminants can simultaneously analyzed in the same reaction for example, in the present work 100 meat samples could have been analyzed in 25 reaction tubes at the same time by using seven primer sets belonging to goat, sheep, pig, cattle, camel, buffalo and donkey. The present study depicts the consequences of an optimized M-PCR, which resulted in a single band of target from one species and no fragment was produced by non-specific amplicon. This study suggests an accurate, sensitive and rapid analytical technique for goat, sheep, pig, cattle, camel, buffalo and donkey meats based on PCR analysis of Cytochrome- b gene of mitochondrial DNA for discovery of meat-adulteration and mixed processed meat.