The highway, which begins at the intersection of Frederica Street and Parrish Avenue in Owensboro, which is also its junction with Kentucky Route 81 and Kentucky Route 2831 (formerly US 431), begins to follow East Parrish Avenue.

With its southeasterly course through Daviess, northeastern Ohio, and northwestern Grayson Counties, KY 54 connects Owensboro to Leitchfield, while also serving the smaller communities of Philpot, Whitesville and Fordsville.

Not only does the road's number equal its rounded mileage, each of the three counties through which it passes (Daviess, Ohio and Grayson) have 18 of the 54 miles.

Initially, KY 54 also ran a course from US 41/60 in downtown Henderson[2] to its present-day western terminus in Owensboro via Hebbardsville[3] and Sorgho,[4] and crossing the Green River via a tolled ferry.

[10] In Owensboro proper, the route has over the years been straightened and widened, particularly toward the eastern reaches of the city.