Rapid Multiplex PCR-Based Identification of Meat Species (SUS SCROFA, EQUUS ASINUS, BOS TAURUS) in Commercial Food Samples
DOI:
https://doi.org/10.57038/usjas.v10i01.7986Keywords:
Simplex PCR, Multiplex PCR, Universal primers, Species-specific primers, Sensitivity, Specificity, Commercial foodstuffsAbstract
For economic, religious, and health-related reasons, it is crucial to safeguard customers against unlawful or undesirable substitutions by evaluating the content and authenticity of food. Red meat is being replaced by the meats of cows, pigs, and donkeys. Because of their religious beliefs, Muslims and Jews abstain from eating pork and donkey meats, even in little amounts. In light of this, multiplex PCR rather than uniplex PCR is required for the quick, affordable and highly sensitive identification of the DNA origins of cow, donkey, and hog species in actual food samples. For this purpose, a total of 85 samples including blood samples, tissue samples, laboratory prepared food samples and commercial foodstuffs including mutton, beef biryani, chicken samosa, chicken pakora, haleem, nihari mutton biryani, etc. were collected from Bahawalpur and Rahim Yar Khan. DNA was extracted from all the targeted samples. In a single tube, we created the first multiplex PCR test to determine the species of cow, donkey and pork used in food items. In addition to universal 18S rRNA primers that amplify a 99 bp fragment, the assay uses specific primers [pork (Sus scrofa), donkey (Equus asinus), and cow (Bos taurus)] that amplify fragments of the mitochondrial COX1 (pork; 460 bp), 12S rRNA (donkey; 184 bp), and ATPase subunit 6 and 8 genes (cow; 271 bp), respectively. For all species, the assay's detection limit was 0.05%. In conclusion, this PCR technique offers an easy, quick, sensitive, accurate, and cost-effective way to identify cow, pork and donkey species in commercial food samples.
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