Background: Extended-spectrum β-lactamases (ESBL) in Escherichia coli are a serious concern due to their role in developing multidrug resistance (MDR) and difficult-to-treat infections. Objective: This study aimed to identify ESBL-carrying E. coli strains from both clinical and environmental sources in Lusaka District, Zambia. Methods: This cross-sectional study included 58 ESBL-producing E. coli strains from hospital inpatients, outpatients, and non-hospital environments. Antimicrobial susceptibility was assessed using the Kirby–Bauer disk diffusion method and the VITEK® 2 Compact System, while genotypic analyses utilised the Illumina NextSeq 2000 sequencing platform. Results: Among the strains isolated strains, phylogroup B2 was the most common, with resistant MLST sequence types including ST131, ST167, ST156, and ST69. ESBL genes such as blaTEM-1B, blaCTX-M,blaOXA-1, blaNDM-5, and blaCMY were identified, with ST131 and ST410 being the most common. ST131 exhibited a high prevalence of blaCTX-M-15 and resistance to fluoroquinolones. Clinical and environmental isolates carried blaNDM-5 (3.4%), with clinical isolates showing a higher risk of carbapenemase resistance genes and the frequent occurrence of blaCTX-M and blaTEM variants, especially blaCTX-M-15 in ST131. Conclusions: This study underscores the public health risks of blaCTX-M-15- and blaNDM-5-carrying E. coli. The strengthening antimicrobial stewardship programmes and the continuous surveillance of AMR in clinical and environmental settings are recommended to mitigate the spread of resistant pathogens.