Abstract Optimising the treatment of respiratory tract infections (RTIs) caused by multidrug-resistant (MDR) Escherichia coli has become challenging with existing antibiotic options. E. coli pathogens have various virulence factors that determine their pathogenesis and antimicrobial resistance (AMR) mechanisms. The evidence of virulence genes harboured by E. coli in human RTIs yet remains unexplored. This study aimed to investigate the presence of fimA, neuC and iutA virulence genes encoding type 1 fimbriae, capsular polysaccharide, and siderophores respectively and AMR pattern of E. coli isolates conferring RTIs. Fifty E. coli isolates were confirmed by culture and biochemical tests. Virulence genes were identified by PCR method. AMR was assessed by disc diffusion method. The prevalence of fimA and neuC was 44%, and 24% respectively though no iutA was found. The frequency distribution of fimA and neuC genes was 42.8% and 25% in upper RTIs and 45.4% and 22.7% in lower RTIs respectively but had no statistically significant difference (fimA, p = 0.854; neuC, p = 0.851) when compared between URTIs and LRTIs. All (100%) E. coli isolates were MDR. Of 50 isolates, highest resistance was observed against amoxicillin (72%) followed by ampicillin (66%), cefixime (56%), sulfamethoxazole/trimethoprim (54%), ciprofloxacin (48%), nalidixic acid (32%) amikacin (28%) and gentamycin (24%). No E. coli including either fimA or neuC positive, was resistant to colistin, imipenem and meropenem antibiotics. This is the first report on virulence-associated genes and MDR among E. coli isolates recovered from RTIs in Bangladesh. Reduced susceptibility of MDR E. coli to most of the antibiotics in RTIs is alarming. Future research should confirm the association between AMR and virulence genes of E. coli isolates identified in RTIs to explain the pathogenesis of RTIs, mechanism of E. coli attaining MDR and antibiotic therapy in future.