Aims To determine if type 2 diabetes mellitus (T2DM) is associated with the spectrum of uropathogens and antimicrobial resistance in urinary tract infections (UTI) in primary care. Methods A cross-sectional study in female outpatients ≥30 years with positive urine cultures. T2DM patients were 1:1 matched to controls by age group and general practitioner (GP). GPs were sent questionnaires for additional data. Uropathogens and resistance patterns were compared between patients with and without T2DM. Multivariable regression analysis was performed to assess the independent association between T2DM and resistance to first line treatments, defined as resistance to nitrofurantoin, trimethoprim, fosfomycin, ciprofloxacin, amoxicillin/clavulanic acid and/or trimethoprim/sulfamethoxazole. Results In 566 urine cultures, 680 uropathogens were found. Resistance to first line treatment antibiotics was present in 62.5% of patients. Frequencies and resistance rates of uropathogens did not differ between both groups of patients. Previous UTI and previous hospital admission were independent risk factors for resistance, but T2DM was not. Conclusions In this study T2DM was not an independent risk factor for antimicrobial resistance in UTI in primary care. Previous UTI and hospitalisation are drivers of resistance and should be included in the decision to perform a urine culture to target first line UTI treatment.