Abstract: Multiple-PCR was conducted to establish a stable PCR system for identifying the three Wx genes in wheat. Two pairs of primers were employed to amplify Wx-A1, Wx-B1, and Wx-D1 genes of wheat, with the target sequences of 230 bp/265 bp, 854 bp, and 204 bp, respectively. The results showed that Wx-A1, Wx-B1, and Wx-D1 can be detected si-multaneously in a single reaction. This method proved to be repeatable and low cost for evaluation of wheat quality proper-ties in breeding program. This multiple-PCR technique can be efficiently used in marker-assisted selection for Wx genes, which will improve selection procedure for waxy wheat.