Antibiotics have increased our lifespan by more than 20 years and shielded us from dangerous microbes. However, their power is dwindling. Therefore, antibiotic resistance is a current worldwide health emergency, and it could be the following pandemic. Thus, the present study assesses the susceptibility and resistance pattern of various E. coli isolates collected from patients at Suez Canal University Specialized Hospital. Collectively, 50 isolates of E. coli out of 150 specimens were isolated from different specimens' types (urine, stool, and blood) and were identified using traditional standard protocols. After performing antibiotic susceptibility testing, the overall tested isolates showed the highest susceptibility against MRP (92%) followed by AK (90%), while the highest resistance was obtained against AMC (100%) followed by CTX (44%). On the other hand, E. coli isolates from urine showed the highest susceptibility against MRP and AK (91.43%) each, while the highest resistance was against AMC (100%) followed by CTX (51.43%). Moreover, the maximum susceptibility of stool isolates was against AK and MRP (83.33 %) each, whereas the isolates obtained from stool showed the highest resistance against AMC (100%) followed by CTX (50%) each. Finally, the E. coli isolates from blood revealed the superior susceptibility to MRP (88.89%) followed by LEV, AK, and DO (77.78%), while the highest resistance of blood specimens was found against AMC (100%) followed by CTX (33.33%). Furthermore, the resistance patterns revealed that 66% of the isolates were MDR. Finally, it is crucial to evaluate the antimicrobial sensitivity test for effective therapy.