The LEE 4 genes sepL, espA, espD, espB, and espF were detected in 50 strains of typical and atypical enteropathogenic (EPEC) and enterohemorrhagic (EHEC) Escherichia coli by PCR. sepL was amplified in 90%, espA in 94%, espB in 50%, espD in 40%, and espF in 78% of all strains, employing prototype EPEC-based primers. With O26:H(-)-based primers, espB was detected in all O26 strains, and O157:H7-specific primers amplified espD and espB among all O55:H7 and O157:H7 strains. Our results indicated that espA and sepL should be more conserved between different EPEC and EHEC serotypes, while espB, espD, and espF should be more diverse. Apparently this variation is related to serogroup or serotype, but sequencing assays are necessary to confirm such conservation/diversity and their association with serogroup or serotype. Secreted protein analyses of espA, espD, and espB PCR-negative strains demonstrated that their encoded proteins present distinct immunological types, reflecting the genetic variability of those genes.