Understanding impacts of disease on wild bird populations requires knowing not only mortality rate following infection, but also the proportion of the population that is infected. Greater sage-grouse (Centrocercus urophasianus) in western North America are known to have a high mortality rate following infection with West Nile virus (WNv), but actual infection rates in wild populations remain unknown. We used rates of WNv-related mortality and seroprevalence from radiomarked females to estimate infection rates in a wild greater sage-grouse population in the Powder River basin (PRB) of Montana and Wyoming from 2003 to 2005. Minimum WNv-related mortality rates ranged from 2.4% to 13.3% among years and maximum possible rates ranged from 8.2% to 28.9%. All live-captured birds in 2003 and 2004 tested seronegative. In spring 2005 and spring 2006, 10.3% and 1.8% respectively, of newly captured females tested seropositive for neutralizing antibodies to WNv. These are the first documented cases of sage-grouse surviving infection with WNv. Low to moderate WNv-related mortality in summer followed by low seroprevalence the following spring in all years indicates that annual infection rates were between 4% and 29%. This suggests that most sage-grouse in the PRB have not yet been exposed and remain susceptible. Impacts of WNv in the PRB in the near future will likely depend more on annual variation in temperature and changes in vector distribution than on the spread of resistance. Until the epizootiology of WNv in sagebrush-steppe ecosystems is better understood, we suggest that management to reduce impacts of WNv focus on eliminating man-made water sources that support breeding mosquitoes known to vector the virus. Our findings also underscore problems with using seroprevalence as a surrogate for infection rate and for identifying competent hosts in highly susceptible species.