To rapidly identify spoilage yeast isolated from food by real-time PCR,several specific probes and primers were designed or selected.The method for identification of Saccharomyces cerevisiae,Zygosaccharomyces rouxii,Dekkera bruxellensis and Spartak pichia by real-time PCR were established in this study.More than 60 strains of yeast isolated from commercial food were identified by real-time PCR method.The results showed that 4 strains of them were Saccharomyces cerevisiae,and 21 strains were Zygosaccharomyces rouxii,the other 35 strains didn't belong to that 4 species of yeast.To identify yeast by real-time PCR method,the whole process took only about 3 hours.Compared with the traditional biochemical method,real-time PCR method was more time-saving,sensitive,and accurate.