This study describes a novel species of Gekko (Squamata: Gekkonidae) based on its distinct morphological features and molecular evidence, which was identified in the Dabie Mountains on the border of Anhui and Henan provinces of Central China. Gekko kaiyai sp. nov. could be distinguished from its congeners owing to its morphological characteristics, such as being a medium body sized gecko species (snout-vent length, 56.98-64.99 mm, n = 4, females; 50.03-61.56 mm, n = 11, males); nostrils scale in contact with rostral scale; tubercles on the dorsal and limb, while the upper forelimb is smooth with no tubercles; 22-33 interorbital scales between the anterior corners of the eyes; 157-209 ventral scales between mental and cloacal slit; 90-121 midbody scale rows; 30-43 ventral scale rows; 7-9 sub-digital lamellae on first fingers, 8-13 fourth fingers, 7-9 first toes, and 7-11 fourth toes; free of webbing in the fingers and toes; 9-12 pre-cloacal pores in males, which are absent in females; post-cloacal unilateral tubercles 1 (few 2); and a dorsum that is greyish white to dark brown, with 6-7 brown markings between the nape and sacrum. The phylogenetic tree based on the mitochondrial DNA sequences (16S, CYTB, and COI) indicated that Gekko kaiyai sp. nov. form an independent clade with strong support (100/1) and are a sister group to G. hokouensis . At the inter-species level, the genetic distances were all large, further confirming that an independent species had been identified. The discovery of this species implies that there are now 87 identified species in the genus Gekko , 22 of which can be found within China.