Resistance to antibiotics among pathogen bacteria is a global concern nowadays. Considering that Escherichia coli (E. coli) is the most important urinary infection and nosocomial infection pathogen, the present study aimed to evaluate the macrolide resistance of E. coli isolated from urinary infection and determine the phylogeny using the ERIC-PCR method. The sampling was performed from patients admitted to Sanandaj Hospitals during 2017. Upon culturing urine samples, the E. coli strains were identified by biochemical tests. Then, the antibiotic sensitivity of the strains was performed using the disk diffusion test. Determining the erythromycin MIC using E-test was performed for species resistant to erythromycin. The bacterial resistance genes were identified using PCR, and ultimately, the gene relationships of the strains were determined using ERIC-PCR. Ninety-nine E. coli strains were isolated from urine samples. The strain resistance to ceftriaxone, piperacillin, erythromycin, azithromycin, carbenicillin, ciprofloxacin, cefotaxime, and amikacin was 51.51%, 84.84%, 84.84%, 30.30%, 75.75%, 41.41%, 84.4%, and 42.42%, respectively. Amon the selected 19 strains, which were resistant to azithromycin and erythromycin, 15 mphA-positive isolates (78.9%), ten mphB-positive isolates (52.63%), and ten isolates (52.63%) simultaneously had mphA and mphB. Also, according to ERICPCR, it was shown that the strains are genetically related, and the same genetic source was not observed between the strains. Considering the prevalence of macrolide-resistant E. coli infections, it suggested that these strains be identified in urinary infection patients. Moreover, treating these patients with macrolide is not recommended. Moreover, the same genetic source was not observed between the strains; therefore, the antibiotic resistance gene spreads in strains with different genetic sources. Thus, there is a need to prevent the overuse of antibiotics.