West Nile virus (WNV) is a public health threat and has caused the death of thousands of North American birds. As such, surveillance for WNV has been ongoing, utilizing numerous biological specimens and testing methods. Nonvascular (i.e., fully grown) feathers would provide a simple method of collection from either dead or live birds of all ages and molt cycles, with presumably less biosafety risk compared with other specimen types, including feather pulp. The current study evaluates WNV detection in nonvascular feathers removed from naturally infected avian carcasses of several species groups. Feathers of corvid passeriforms had the highest sensitivity of detection (64%), followed by noncorvid passeriforms (43%), columbiforms (33%), and falconiforms (31%). Storing feathers for 1 year at -20 degrees C or at ambient room temperature resulted in detection rates of infectious WNV of 16% and zero, respectively, but had no effect on detection rates of WNV RNA in a subset of matched feather pairs (47% for both storage temperatures). The efficacy of WNV detection in nonvascular feathers is greatly enhanced by testing multiple feathers. The advantages of using nonvascular feathers over other tissues may outweigh the relatively low detectability of WNV RNA in certain situations such as remote areas lacking resources for acquiring other types of samples or maintaining the cold chain.