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Abstract Our study was carried out to isolate E.coli from foods from different sourcesA total of 360 samples were collected from different areas in Ismailia, Egypt, the samples collected were 100 minced meat, 100 raw milk samples, 100 Kareesh cheese and 60 diarrheal stool samples and analyzed using cultural methods, 115 E.coli isolates were detected and identified by biochemical tests (IMVC) and serologically. Among the E.coli isolates the percentages were 46.6% isolates from human stools samples, 35% from meat samples, 31% from kareesh cheese and 21% from raw milk samples. Serological identification revealed that 24.3% of E.coli isolates were typable and 75.7% were untypable. The highest percentage was EPEC and EHEC (8.7%), (7%) respectively followed by EIEC 5.22%, ETEC were 4.35% and the least % was EAEC 2.61%. Multiplex PCR technique for detection of stx1, stx2, eaeA and EHEC hlyA genes. Multiplex PCR technique was conducted on all E.coli isolates, we identified 10 E.coli isolates positive for one gene or more for the four tested genes (Stx1, Stx2, eaeA, hlyA). 8 isolates possessed intimin gene (eaeA) from all the four sources of sample collection (3 from meat, 2 from raw milk, 1 from cheese and 2 from children); 2 isolates carried Stx2 (1 from children and 1 from cheese) ; Stx1 was only identified in 1 isolate isolated from milk. The antibiotic sensitivity pattern showed that the identified serotypes were highly sensitive to amikacin (100%), ciprofloxacin and chloramphenicol (96.5%), followed by Cefatriaxone and azithromycin (93%), and there were resistance to erythromycin(71.5%) andstreptomycin(46.4%) and to sulphamethoxazole/ trimethoprim (14.2%). Furthermore, the present study failed to isolate E.coli O157 from any of the examined samples |