Cambarus (Puncticambarus) brimleyorum is a new species of crayfish currently known only from lotic habitats in the Hiwassee River basin in Cherokee and Clay counties, North Carolina. Morphologically and meristically it is similar to another endemic Hiwassee crayfish, C. (P.) parrishi Hobbs, 1981, from which it differs in having a rostrum that lacks marginal spines or tubercles, a dorsal surface that is usually subplane instead of excavate, and a consistently longer acumen. The two species also vary in the shape of the cephalic lobe of the epistome. The new species differs from the only other species of Puncticambarus that occurs in the Hiwassee River basin, C. (P.) hiwasseensis Hobbs, 1981, in lacking rows of prominent tubercles on the mesial surface of the dactyl of the chela of the cheliped, in its narrower, less punctate areola, and in a number of other characters. At some sites, C. brimleyorum is syntopic with either C. hiwasseensis or C. parrishi, but these latter species have never been found together, and C. parrishi is still known only from a few headwater tributaries in Clay County, North Carolina, and Towns County, Georgia.