Abstract As a result of the COVID-19 pandemic, HIV care and prevention efforts have been disrupted. We investigated pre-exposure prophylaxis (PrEP) use and testing behaviors among MSM in the Netherlands, and the factors that influenced testing behaviors during the COVID-19 pandemic. A cohort of 766 MSM, established in 2017, was asked in August 2020 to report on their experiences during the COVID-19 pandemic via an online survey. Participants (n = 319) reported increased PrEP use and, among PrEP users (n = 211), significantly lower rates of having tested in the last 3 months for HIV and renal functioning compared to before the pandemic. Daily PrEP use and a higher number of sexual partners during the pandemic was significantly associated with continued HIV testing. Continued renal functioning testing was associated with older age. Correcting for pandemic-related disruptions in PrEP use and care will require sustained effort to understand and address missed opportunities.