The event of pathogenic microorganisms in ecological waters is a progressing worry for general wellbeing authorities and those in the water administration territory around the world. Escherichia coli (E. coli) is considered as a standout amongst the most vital reasons for bacterial diseases transmits over foods and water. The pathogenecity relies upon the outflow of a variety of destructiveness elements created via E. coli. The point of this examination was to explore the prevalence of E. coli strains carrying virulence genes eltB, PCVD, VT1, VT2, ial, isolated from different water sources in Alborz province, Iran. The research was conducted from September 2013 to September 2014. The study included all E. coli strains isolated from different surface water sources (surface water, refinery inputs, water well) in Alborz province of Iran. E. coli isolates were detected and identified via biochemical tests and standard microbiological. The strains were evaluated for the occurrence of virulenc genes eltB, PCVD, VT1, VT2, ial, by PCR using specific primers. The PCR amplicons were visualized via electrophoresis and stained by ethidium bromide. One hundred E. coli strains were isolated and involved in the research. The frequency of virulence genes was: VT2 (43%), eltB (19%), Ial (15%), PCVD (13%), VT1 (3%). Our finding showed the prevalence rate of virulence genes VT2 is very high among E. coli strains isolated from different surface water sources in Alborz province. Considering their plasmid borne nature, the risk of transmission of these genes between other bacterial species could pose a high threat for public health.