According to this result our method was successful in the differentiation of the three Entamoeba species. Further, to ensure that the presence of other Entamoeba species that could be found colonizing the human intestine does not interfere with the interpretation of restriction patterns, a second analysis was conducted in silico including sequences of E. coli, E. hartmanni and E. polecki. This analysis showed that all six species produced different restriction patterns readily identifiable, as seen in Fig. 3.