Estimation of forest cover change is important for boreal forests, one of the most extensive forested biomes, due to its unique role in global timber stock, carbon sequestration and deposition, and high vulnerability to the effects of global climate change. We used time-series data from the MODerate Resolution Imaging Spectroradiometer (MODIS) to produce annual forest cover loss hotspot maps. These maps were used to assign all blocks (18.5 by 18.5 km) partitioning the boreal biome into strata of high, medium and low likelihood of forest cover loss. A stratified random sample of 118 blocks was interpreted for forest cover and forest cover loss using high spatial resolution Landsat imagery from 2000 and 2005. Area of forest cover gross loss from 2000 to 2005 within the boreal biome is estimated to be 1.63% (standard error 0.10%) of the total biome area, and represents a 4.02% reduction in year 2000 forest cover. The proportion of identified forest cover loss relative to regional forest area is much higher in North America than in Eurasia (5.63% to 3.00%). Of the total forest cover loss identified, 58.9% is attributable to wildfires. The MODIS pan-boreal change hotspot estimates reveal significant increases in forest cover loss due to wildfires in 2002 and 2003, with 2003 being the peak year of loss within the 5-year study period. Overall, the precision of the aggregate forest cover loss estimates derived from the Landsat data and the value of the MODIS-derived map displaying the spatial and temporal patterns of forest loss demonstrate the efficacy of this protocol for operational, cost-effective, and timely biome-wide monitoring of gross forest cover loss.