Object： To establish a method for rapid detection of diarrhea-causing Escherichia coli（DEC）. Methods： Primers and probes were designed based on uidA（GenBank： JQ068101.1）， bfpB （Sequence： NG_034726.1）， eaeA （GenBank： AJ579305. 1）， LT （GenBank： M17873.1）， ST （GenBank： M18346.1）， Stx1 （GenBank： FR875155.1）， Stx2 （GenBank： AB030484.1）， ipaH （GenBank： JQ638638.1）， aggR （GenBank： Z32523.1） of DEC to establish a TaqMan probe fluorescent PCR assay. Among these targets， uidA gene was used to identify all DEC， and the remaining targets are divided into two systems， A and B. A system including 4 genes（bfpB， eaeA， LT， ST） were used to detect EPEC and ETEC， B system includes 4 genes（ipaH， Stx1， Stx2， and aggR） were used to detect EIEC， STEC and EAEC. The 5' ends of the two system probes were labeled with FAM， VIC， TET and ROX respectively. Results： The amplification efficiency of the two systems was 89.6%-102.2% respectively and the linear correlation coefficient R2 was in the range of 0.965-0.999. This assay is specific to Escherichia coli（25/25）， and the Real-time PCR detection showed that EPEC， ETEC， EIEC， STEC and EAEC could be identified and distinguished by system A and B while 70 non-DEC amplification results were negative. Conclusion： This new method is highly specific to E coli and can accurately distinguish EPEC， ETEC， EIEC， STEC and EAEC.