Gold mining in Amazonia involves forest removal, soil excavation, and the use of liquid mercury, which together pose a major threat to biodiversity, water quality, forest carbon stocks, and human health. Within the global biodiversity hotspot of Madre de Dios, Peru, gold mining has continued despite numerous 2012 government decrees and enforcement actions against it. Mining is now also thought to have entered federally protected areas, but the rates of miner encroachment are unknown. Here, we utilize high-resolution remote sensing to assess annual changes in gold mining extent from 1999 to 2016 throughout the Madre de Dios region, including the high-diversity Tambopata National Reserve and buffer zone. Regionally, gold mining-related losses of forest averaged 4437 ha yr-1. A temporary downward inflection in the annual growth rate of mining-related forest loss following 2012 government action was followed by a near doubling of the deforestation rate from mining in 2013-2014. The total estimated area of gold mining throughout the region increased about 40% between 2012 and 2016, including in the Tambopata National Reserve. Our results reveal an urgent need for more socio-environmental effort and law enforcement action to combat illegal gold mining in the Peruvian Amazon.