To understand the distribution of diarrheagenic Escherichia (E.) coli in population in Shanghai and discuss the practice model of cooperation in enteric infectious disease prevention and control between public health institution and hospital.Sentinel hospitals were assigned, standard detection and identification of diarrheagenic E. coli were conducted, incidence curve of diarrheagenic E. coli infection was drawn and epidemiologic survey and laboratory detection were conducted for suspect diarrheagenic E. coli infection outbreaks.A total of 7 204 stool specimens were collected from diarrhea patients in 4 hospitals during 2012-2013, in which 712 (9.9% ) were diarrheagenic E. coli positive, including 351 enteropathogenic E. coli (EPEC) strains, 292 enterotoxigenic E. coli (ETEC) strains, 32 enteroinvasive E. coli(EIEC) strains and 6 Shiga toxin-producing E. coli (STEC/EHEC) strains, as well as 31 mixed strains. EPEC infection mainly occurred in children aged 1-5 years; and all of these infections were caused by aEPEC. The incidence peak of ETEC infection was during August, the positive rate was >20%. The ETEC infection mainly occurred in infants aged 1-28 days in 2012 and in people aged 20-60 years in 2013 (P<0.05). ST was the major type (59.6%), followed by LT (27.8%) and ST/LT (12.6%). EIEC infection increased in children obviously in 2013 (P<0.01). No EHEC O157:H7 case was detected, but two EHEC O26:H11 (eae-hlyA-stx1a) cases in children were reported for the first time in Shanghai. The survey result indicated that the multidrug-resistant ETEC (STh-CS21-CFA/I-ClyA-EatA-ST2332-SHNL0005) strain causing outbreak in 15 newborns in Shanghai in 2012 was in the same clone as the strain detected in Zigong in Sichuan province.Significant change has occurred in diarrheagenic E. coli distribution in Shanghai in recent years, ETEC has potential risk to cause outbreak of hospital acquired infection in neonates and food borne infection. The active surveillance on ETEC and other enteric pathogens by both public health institutions and hospitals need to be improved.