For the first time, here we described a multiplex polymerase chain reaction assay for the accurate identification of five
meat species forbidden in Islamic foods in a single assay platform. Five pairs of species-specific primers
were designed targeting mitochondrial ND5, ATPase 6, and cytochrome b genes to amplify 172, 163, 141,
129 and 108 bp DNA fragments from cat, dog, pig, monkey and rat meats, respectively.All PCR products
were identified in gel-images and electrochromatograms obtained from Experion Bioanalyzer. Species-specificity
checking against 15 important meat and fish and 5 plant species detected no cross-species
amplification. Screening of target species in model and commercial meatballs reflected its application
to detect target species in process foods. The assay was tested to detect 0.01–0.02 ng DNA under raw
states and 1% suspected meats in meatball formulation.