Eliminating HIV transmission and therefore ending the HIV epidemic is a feasible goal for 2030, if interventions to prevent HIV transmission are optimally applied. Diagnosing individuals with HIV is one of the key prerequisites to prevent HIV transmission. Additionally, timely diagnosis and treatment of HIV improves patient’s health outcomes. Healthcare providers in the primary care and hospital setting may contribute by implementing proactive, provider-initiated testing strategies. We compared HIV testing by general practitioners versus sexual health centers in five Dutch regions and concluded that general practitioners play a significant role in HIV testing and diagnosis. Additionally, we assessed the implementation of indicator condition-guided testing in various healthcare settings and found that there is a large variation in its implementation, with overall low HIV test ratios. We implemented multifaceted educational interventions in the primary care and hospital setting to improve providers’ HIV testing strategies. These interventions yielded modest but statistically significant increases in HIV testing. Lessons learned on determinants for HIV testing as well as identified persistent barriers and HIV-related stigma among patients and healthcare providers should be addressed in our efforts to improve HIV testing in healthcare settings moving forward. Ultimately, this will contribute to a reduction of the proportion of people with HIV who remain undiagnosed and the proportion diagnosed at a late-stage HIV infection, thereby improving individual’s health outcomes, and preventing onward transmission of HIV.