A rapid and low-cost diagnosis for EPEC/EHEC infections is extremely required considering their global prevalence, the severity of the diseases associated with them, and the fact that the use of antibiotics to treat EHEC infections can be harmful. For EHEC, the detection of Stx toxins has already been developed, but for EPEC, an internationally recognized standard diagnostic test is lacking. Thus, the approach for their rapid detection in this study was the use of the secreted proteins EspA and/or EspB, since they are the major secreted proteins in both pathogens. EspB was defined as a biomarker and its corresponding monoclonal antibody as the tool for EPEC/EHEC diagnosis using a latex agglutination assay, which can be employed in less equipped laboratories in developing countries.