Background; Human immunodeficiency virus (HIV) and hepatitis C (HCV) screening and human papillomavirus (HPV) vaccine uptake remain suboptimal. To improve HIV and HCV screening and HPV vaccination, we implemented a quality improvement project in three southwestern Pennsylvania family medicine residency practices. Methods: From June 1 to November 30, 2021, participating practices used universal screening and vaccination guidelines and chose from multiple strategies at the office (e.g., standing orders), provider (e.g., multiple forms of provider reminders), and patient (e.g., incentives) levels derived from published literature and tailored to local context. Age-eligible patients for each recommendation with at least one in-person office visit during the intervention period were included. To assess the interventions' effect, we obtained testing and vaccination data from the electronic health record for the intervention period, contrasted it with identical data from June 1 to November 30, 2020, and used logistic regression controlling for patient age, sex, and race to determine differences in screening and vaccination between intervention and baseline periods. Results: A total of 14,920 and 15,523 patients were eligible in the baseline and intervention periods, respectively. Following the intervention, HIV lifetime screening but not first-time screening for patients 13 to 64 years old was significantly higher (78.9% vs. 76.1%, p = 0.004 and 39.6% vs. 36.6%, p = 0.152, respectively; adjusted odds ratio for increase [aOR] = 1.21; 95% confidence interval [CI] = 1.06, 1.38). HCV lifetime screening for patients 18 to 79 years old was significantly higher post-intervention (62.5% vs. 53.5%, p < 0.001; aOR = 1.51; 95% CI = 1.4, 1.64). For patients 9 to 26 years old, no change in HPV initiation was observed but the percentage of patients who completed their HPV vaccinations in the observed period was significantly higher post-intervention (4.6% vs. 7.0%, p = 0.006, aOR = 1.58; 95% CI = 1.14, 2.2). During the post-intervention period, we identified 0 new HIV diagnoses and 48 HCV diagnoses (19 eligible for treatment). Conclusions: Family medicine residency office-based multi-strategy efforts appear to successfully increase patient uptake of HIV and HCV screenings and maintain HPV vaccination rates.