Previous (1992–1994) surveys for native freshwater mussels (Unionidae) along main channels of the Detroit River showed that unionids had been extirpated from all but four sites in the upper reaches of the river due to impacts of dreissenid mussels (Dreissena polymorpha and D. bugensis). These four sites were surveyed again in 1998 using the same sampling method (timed-random searches) to determine if they may serve as “refugia” where unionids and dreissenids co-exist. Two additional sites were sampled using additional methods (excavated-quadrat and line-transect searches) for comparison with unpublished data collected in 1987 and 1990. A total of four individuals of four species (Actinonaias ligamentina, Cyclonaias tuberculata, Lasmigona complanata and Pleurobema sintoxia) were found by timed-random searches at four sites in 1998 compared to 720 individuals of 24 species in 1992 and 39 individuals of 13 species in 1994. Excavated-quadrat and line-transect searches at the two additional sites yielded only one live specimen of Ptychobranchus fasciolaris compared to 288 individuals of 18 species in 1987 and 1990. Results of this study suggest that remaining densities of unionids in channels of the Detroit River are too low to support viable reproducing populations of any species. Therefore, we conclude that unionids have been extirpated from main channels of the Detroit River due to dreissenid infestation. As the Detroit River was one of the first water bodies in North America to be invaded by dreissenids, it is likely that unionids will also be extirpated from many other rivers and lakes across eastern North America over the next few decades. Resource agencies should be encouraged to implement active management programs to protect remaining unionid populations from zebra mussels.