Abstract: A PCR for detection of two categories of diarrheagenic Escherichia coli was developed. This method proved to be specific and rapid in detecting virulence genes from enteropathogenic E. coli (EPEC) (eae and bfp) and Shiga toxin-producing E. coli (STEC) (stx1, stx2 and eae) from seventy isolates of various sources. Present results confirm that it is possible and feasible to perform a simultaneous amplification of the virulence genes from two categories of diarrheagenic E. coli (STEC, EPEC) and that this technique becoming a novel diagnostic tool for future water food-borne outbreaks studies.