Urinary tract infection (UTI) is a common cause for prescription of antibiotics among women in general practice. Diagnosis is often established by inquiry into clinical history and symptoms, and these may be experienced differently depending on menopause status of the woman. The aim of this study was to assess differences in severity and bothersomeness of UTI symptoms between pre- and postmenopausal women. We used a convenience sample of 313 women with suspected UTIs and typical symptoms recruited in general practice. Each woman completed the Holm and Cordoba UTI score (HCUTI), measuring the severity and bothersomeness of the dimensions: dysuria, frequency, lower back, and general symptoms. The exposure was menopausal status. Differences in the various HCUTI dimensions between the menopause groups were investigated in linear regression models, adjusting for potential confounders. Premenopausal women had a significantly higher severity score for the item "feeling unwell" than postmenopausal women (mean difference -0.59, 95% CI -0.88 to -0.31). They also had a significantly higher bothersomeness score for the items "pain on urination" (mean difference -0.54, 95% CI -0.83 to -0.25), "feeling unwell" (mean difference -0.62, 95% CI -0.92 to -0.32), and for the dimension "dysuria" (mean difference -0.38, 95% CI -0.61 to -0.15) than postmenopausal women. This study found differences in some aspects of symptom severity and bothersomeness between pre- and postmenopausal women presenting in general practice with suspected UTIs. Menopausal status should be taken into account when using symptoms to diagnose and evaluate response to UTI treatment in both clinical practice and research.