We describe a new species of the gekkonid genus Cyrtodactylus from Khammouane Province, central Laos based on morphological and molecular data. Morphologically, Cyrtodactylus rufford sp. nov. differs from its congeners by a unique combination of the following characters: medium size, SVL reaching 72.5 mm; dorsal pattern with three or four light transverse bands between limb insertions; one intersupranasal; 14-16 irregular dorsal tubercle rows at midbody, weakly developed in the paravertebral region; 27-29 ventral scale rows between ventrolateral folds; 42-43 precloacal and femoral pores in a continuous row in males, enlarged femoral and precloacal scales present; 4 or 5 postcloacal tubercles on each side; dorsal tubercles present at base of tail; medial subcaudal scales enlarged. Molecular analyses show that the new species is closely related to C. khammouanensis, which was originally described from Khammouane Province.