Tripylina puxianensis n. sp. is described and illustrated from Shanxi province, China. It is characterized by having a triangular dorsal tooth, pointing towards the ventral side, anterior to two sub-ventral teeth, with dorsal and sub-ventral teeth similar in shape and size; one ventromedian seta in the cervical region; vulva with protruding lips, absence of post-uterine sac; and a pair of setae on the tail; male not found; female body length 1048-1331 µm, a = 25.5-33.8, b = 5.5-6.2, c = 14.4-23.1, c' = 2.1-3.5 and V = 56.1-68.5%. The phylogenetic relationships among species in the genus Tripylina were analyzed using data from the SSU (small subunit) and D2/D3 expansion segments of the LSU (large subunit) rDNA. These analyses confirmed that T. puxianensis n. sp. is different from other members of the genus for which sequences are available.