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We examined the prevalence trends of non-human immunodeficiency virus (HIV) sexually transmitted infections (STI) and associated patient characteristics in U.S. ambulatory-care settings from 2005–2016. We conducted a retrospective repeated cross-sectional analysis using data from the National Ambulatory Medical Care Survey (NAMCS) for individuals aged 15–64 with a non-HIV STI-related visit. Data were combined into three periods (2005–2008, 2009–2012, and 2013–2016) to obtain reliable estimates. Logistic regression was used for analysis. A total of 19.5 million weighted, non-HIV STI-related ambulatory visits from 2005–2016 were identified. STI-related visits per 100,000 ambulatory care visits increased significantly over the study period: 206 (95% CI = 153–259), 343 (95% CI = 279–407), and 361 (95% CI = 277–446) in 2005–2008, 2009–2012, and 2013–2016, respectively (Ptrend = 0.003). These increases were mainly driven by increases in HPV-related visits (56 to 163 per 100,000 visits) from 2005–2008 to 2009–2012, followed by syphilis- or gonorrhea-related visits (30 to 67 per 100,000 visits) from 2009–2012 to 2013–2016. Higher odds of having STI-related visit were associated with younger age (aged 15–24: aOR = 4.45; 95% CI = 3.19–6.20 and aged 25–44: aOR = 3.59; 95% CI = 2.71–4.77) vs. 45–64-year-olds, Black race (aOR = 2.41; 95% CI = 1.78–3.25) vs. White, and HIV diagnosis (aOR = 10.60; 95% CI = 5.50–20.27) vs. no HIV diagnosis. STI-related office visits increased by over 75% from 2005–2016, and were largely driven by HPV-related STIs and syphilis- or gonorrhea-related STIs. [ABSTRACT FROM AUTHOR]