In spite of the role of integrons as the main contributor to multidrug resistance worldwide, their prevalence in Egypt is still underestimated. In this work, we announce the emergence of class 2 and 3 integrons among Enterobacteriacae isolates from Mansoura University Hospitals. Ninety-three clinical isolates were obtained from different clinical sources, among which 70% of E. coli, 94.8% of K. pneumoniae and 85.7% of Enterobacter spp. were assigned to be multidrug resistant (MDR). Subsequently, the occurrence of class 1-3 integrons was confirmed by multiplex PCR. Class 1 integron was the most predominant being harbored by 42.8%, 90% and 25% of MDR E. coli, K. pneumoniae and Enterobacter spp. isolates, respectively. This was followed by class 2 and 3 integrons which were, for the first time, reported in these hospitals. Also, coexistence of integrons 1and 2 was revealed in 36.9% of integron positive isolates. A significant association was noticed only between resistance to gentamicin and integron prevalence among MDR E. coli isolates (P = 0.02). In conclusion, this work represents the first report for detection of class 2 and 3 integrons, beside the previously detected class 1 integrons. This highlights the high incidence of integrons among MDR Enterobacteriacae isolates which indicates the selective pressure of antibiotics in these hospitals. Moreover, this study confirms the possibility of the use of integrons as markers for MDR identification.