To distinguish sika deer (Cervus nippon) tissue samples from those of sympatric cervids in the southern part of Anhui Province, China, we analysed randomly amplified polymorphic DNA (RAPD) fragments of 12 individual samples from four cervid species: black muntjac (Muntiacus crinifrons), Reeve’s muntjac (Muntiacus reevesi), sika deer (Cervus nippon), and tufted deer (Elaphodus cephalophus). Only one primer among the 100 screened produced a clear specific band for identifying DNA of sika deer. We cloned and sequenced 449 bp from this fragment of DNA. We then designed a pair of 18 bp primers (MHL-U/MHL-D) according to the sequence, resulting in a 251 bp sequence-characterised amplified region (SCAR) for sika deer. By combining this pair of SCAR primers with a universal set of mammalian DNA primers, the identification of sika deer tissue samples by a simple common multiplex PCR assay is straightforward, rapid, and reliable. The method will be useful for cervid conservation and cervid bushmeat trade regulation.