16S RRNA PCR-RFLP FOR SPECIATION OF MEAT OF PORCINE ORIGIN IN PROCESSED AND ADMIXED MEATS

摘要

The Polymerase Chain Reaction (PCR) - Restriction Fragment Length Polymorphism (RFLP) using 16S Ribosomal ribonucleic acid (rRNA) gene was explored to test its effectiveness in the identification ofpigmeat in raw, cooked and admixed meat Critical perusal of the restriction maps of ~59Tbase pairs (bp) 16S Ribonucleic acid (RNA) fragment sequences from pig as well as other livestock identified Rsal restriction enzyme (Rsal) specific to pig, yielding 343 bp and 251 bp fragments in the pig. In other livestock species such as cattle, buffalo, goat and sheep, no restriction enzyme site was found. In all the samples of raw pork, the restriction digestion of594 bp fragment with Rsal yielded the 343 bp and 25 7 bp fragments, with no restriction digestion inall other livestock species i. e. cattle, buffalo, goat and sheep. PCR-RFLP with Rsal of594 bp fragment was amplified from three different types of cooked meat DNA samples from pig i. e. cooking at 72°Cfor 30 min, steam cooking at 90°C for 30 min and autoclaving at 120°C/15 lb/30 min and yielded similar restriction enzyme digestion profile as for the raw meat Deoxyribonucleic acid (DNA). This suggested no effect of cooking on PCR-RFLP. Similarly, PCR-RFLP was able to differentiate the origin of meatin mixed meat samples quite consistently up to the level of 25%. However, at levels below 10% results were not consistent.