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Abstract In the present investigation, some phenotyic and molecular typing techniques were applied on 108 strains of E. coli, 88 strains of Ps. aeruginosa and 8 strains of Serratia isolated from Mansoura University Hospitals from different clinical lesions. The distribution of antibiotic resistance among the isolated strains showed high incidence of resistance and imipenem was the most active antibiotic. Using the active pyocin typing, 72 strains of Ps. aeruginosa could be typed into 35 pyotypes. SDS-PAGE of total cell protein extracts showed that the presence of fifteen patterns among E. coli strains and eleven patterns among Ps. aeruginosa strains. Using PCR technique it was found that 84% of the 50 tested strains were found to have at least one of the tested ESBLs also TolC and AcrA genes were present in all tested E. coli except 4 strains and did not present in Ps. aeruginosa except 4 strains. Plasmid profiles of 23 tested E. coli appear to be diverse also the prevalence of plasmids in 22 tested Ps. aeruginosa strains was low than in tested E. coli. Using Pyrosequencing technique, the sequenced region of gyrA, gyrB and ParC were able to differentiate between the tested strains. And in the present study, molecular cloning of the whole sequence of bla-TEM, bla-SHV and bla-CTX-M was carried out experimentally to study the expression of these genes and determine which genes of them responsible for the resistance. |