The subfamily Murininae has high species diversity in Vietnam, but taxonomic studies are limited. In this paper, we describe a new species of the genus Murina based on a specimen collected from Ngoc Linh Nature Reserve, Kon Tum Province in the Central Highlands of Vietnam. It is a medium-sized species with ‘suilla-type’ dentition. A taxonomic review of Murininae from Vietnam was also conducted based on combined morphological, DNA, and karyological characteristics. Molecular phylogenetic analyses based on the mitochondrial cytochrome c oxidase subunit (COI) gene supported the subfamily Murininae, while the genus Murina proved to be paraphyletic in relation to the genera Harpiocephalus and Harpiola. Fourteen species of the genus Murina, one species of Harpiocephalus, and one species of Harpiola are recognized from Vietnam. Murina tiensa is regarded as a junior synonym of M. harrisoni; strong sexual dimorphism was observed in M. harrisoni. Relations between forearm length and total length of skull showed different trends among species and sexes. Karyotypes of Murina huttoni, M. cyclotis, M. lorelieae, M. beelzebub, M. feae, and Harpiola isodon were 2n = 44, FN = 50, while that of Harpiocephalus harpia was 2n = 44, FN = 52.