Multidrug-resistant Escherichia coli have become a main public health distress in Pakistan and many countries, causing failure in treatment with the resultingenormous health burden. The current study was aimed to find the prevalence of E. coli among clinical isolates and their antibiotics susceptibility pattern using standard procedures. The Purpose of the present study to investigate the Prevalence and antibiotic susceptibility pattern of E. coli detected from clinical patients visiting Khyber teaching hospital Peshawar. In this study a total 415 sample were isolated from the patient of Khyber teaching hospital (KTH) Peshawar. The collected samples were processed for identification including Gram staining and biochemical test. Furthermore, the antibiotic susceptibility pattern of all the positive strains of E. coli was tested using selected antibiotic discs by disc diffusion method as per CLSI guidelines 2019.A total of 415 samples, 112 clinical isolates yielded the growth of E. coli using standard procedures. Out of 112 isolates, 52% were recovered from male patients while 60% were females. The positive samples were obtained from urine (58%), pus (14.3%), swab (8.9%), sputum (10.7%), and others 8.0% respectively. The highest sensitive drugs are FOS (99.1%), AK (98.2%), TZP (97.3%), MEM (93.8%), TGC (91.1%), CN (89.3%), SCF (78.6%), DO (65.2%), CT (60.7%), F (49.1%), ATM (27.7%) AMP (23.2%), CAZ (23.2%), CPM (22.3%) , CRO (19.6%) ,CTX (13.4%) and the lowest sensitive drug is CIP which is (7.1%) . The drugs which show high resistance are CIP (92.9%), CTX (86.6%), CRO (80.4%), CPM (77.7%), CAZ (76.8%), AMP (76.8%) , ATM (72.3%) , CT( 39.3%) , DO (34.8%), SCF (21.4%), CN(10.7%),TGC(8.9%), F(8.9%),MEM(6.3%) ,TZP(2.7%),AK (1.8%) ,and the lowest resistance drug is FOS (0.9%).Drug resistance monitoring and the epidemiological analysis of patient data are needed regularly and can be useful for the adequate management of antimicrobial resistance.