Epidemiological studies suggest frequent association of enteropathogenic bacteria with Entamoeba histolytica during symptomatic infection. In this study, we sought to determine virulence gene of enteropathogenic E. coli accompanied to the infection of Entamoeba histolytica. A total of 100 samples (stool and blood) were collected from patients suffering from diarrhea with inflammatory feature from several different locations in the AlDiwaniyah City, Iraq, including: Al-Diwaniya Teaching Hospital, Maternityand Children Hospital, Hamza general Hospital and some many private medical laboratory during the period from october2021 to April 2022. The current study was done at AL-Qadisiyah University. The stool samples were classified to healthy, infected with E.histolytica according to general stool examination, then morphological and genetic examinations were done. The results showed from total samples 12% with E.histolytica infection without bacteria, 48%with parasite infection accompanied by bacteria and 30% within bacterial infection only. According to age category the results showed the category 15-60 It showed the highest rate of infectionsamong the other groups. The higher percent infection was E Coli from a total of 4 samples the infection was 25. bacteria were diagnosis using PCR 16s E Coli showed 25 from total positive for bacterial infections (40). The virulence factor showed in E. Coli the HlyA gene appeared in 20 samples,Klebsiella spp. showed blaCTX-Min 7 samples.The study concluded that the most common bacterial species associated with Entamoeba histolytica is E. coli, and the most virulence factor associated with this infection is the HlyA gene

E. histolytica, HlyA gene, E.coli,blaCTX-M