||Wheat (Triticum aestivum L.) quality properties are strongly affected by the compositions of high-molecular-weight glutenin subunits, kernel hardness, amylase content, pre-harvest sprouting tolerance and presence or absence of 1B/1R translocation. It is very important to develop multiplex PCR for wheat quality improvement in molecular marker assisted breeding to reduce the cost and improve the efficiency. Three types of multiplex PCRs were developed and validated with 13, 30, and 11 Chinese wheat cultivars and advanced lines with known genes, respectively. The first multiplex PCR was used to simultaneously detect genes ω-secalin (1B/1R), Vp1B3, and Pinb-D1b for improving wheat processing quality. The second one was to detect the genes ω-secalin, Ax2*, Bx17, and Dx5 for improving gluten quality and bread making quality. The third multiplex PCR included three markers for Wx-7A, Wx-4A, and Wx-7D to improve starch quality and breed waxy wheat cultivars. The genotypes of all tested wheat cultivars and advanced lines identified by three multiplex PCRs were in agreement with those detected by other methods. The primer-primer interactions in each multiplex PCR were not found. Genomic DNAs extracted from both wheat kernels and leaves were feasible for three multiplex PCR amplifications. The three multiplex PCRs were highly effective in the test of Chinese wheat cultivars, demonstrating good repeatability and low cost for the evaluation of wheat quality properties in wheat breeding program.