The present invention discloses a multiplex polymerase chain reaction (Multiplex PCR) kit for multiple target detection in a single assay platform for Halal authentication. The Multiplex PCR kit disclosed according to the present invention comprises of five sets of forward and reverse specifically designed primers for DNA amplification to identify five meat species forbidden in Islamic foods such as pig, dog, cat, rat and monkey. In another embodiment, the multiplex PCR kit according to the present invention comprising five different sets of species-specific primers that specifically detect five different segments (108 to 172 bp) of three different mitochondrial genes of the said five species.