The present invention discloses a kit for detecting multiple targets in a food, said kit comprising: a set of primer pair for each of the targets for detecting the target DNA; wherein the target DNA are mitochondrial genes including cytochrome b and NADH dehydrogenase 5; characterized in that: said multiple targets comprising cow, buffalo, chicken, duck, goat, sheep and pig; the primer pair of cow comprises a forward primer with SEQ ID NO: 1 and a reverse primer with SEQ ID NO: 2; the primer pair of buffalo comprises a forward primer with SEQ ID NO: 3 and a reverse primer with SEQ ID NO: 4; the primer pair of chicken comprises a forward primer with SEQ ID NO: 5 and a reverse primer with SEQ ID NO: 6; the primer pair of duck comprises a forward primer with SEQ ID NO: 7 and a reverse primer with SEQ ID NO: 8; the primer pair of goat comprises a forward primer with SEQ ID NO: 9 and a reverse primer with SEQ ID NO: 10; the primer pair of sheep comprises a forward primer with SEQ ID NO: 11 and a reverse primer with SEQ ID NO: 12; and the primer pair of pig comprises a forward primer with SEQ ID NO: 13 and a reverse primer with SEQ ID NO: 14; wherein the primer pairs of SEQ ID No: 1 to SEQ ID NO: 14 detect target DNA fragments in a range of 73 base pair to 263 base pair.