A rapid and accurate Polymerase Chain Reaction (PCR) method for detection of Proteus.spp was established in this study.Proteus genus-specific primers were designed based on the sequence polymorphism of atpD gene.The PCR method was validated using a total of 66 Proteus strains including type strains and clinical isolates,and was compared with two culture-based methods:plate-culture method and Vitek Auto Microbe System.Results showed that the detection time of this PCR method was only 5-6 h,which is almost 40 h less than the conventional plate-culture method.Further sensitivity and specificity tests indicated that the PCR method is superior to the plate-culture method and Vitek Auto Microbe System in the detection of Proteus strains.