Vitronura setaebarbulata sp. nov. and V. quartadecima sp. nov. are described from Shanghai, China. V. setaebarbulata sp. nov. is characterized by its heavy barbulate macrosetae, very granulated tubercles, dorsointernal tubercles on head coalescent and very close each other, mandible with three big teeth, apical one with denticules and maxilla with one of the 2 lamellae trifurcated. V. quartadecima sp. nov. is characterized by having 14 tubercles on head, well isolated each other, smooth body setae and mandible with three big teeth, apical one with denticules. The COI barcode sequences were sequenced for four individuals of V. setaebarbulata sp. nov. and twelve individuals of V. quartadecima sp. nov. respectively. No genetic divergence between individuals of same species was found, and the divergence of nucleotides between the two new species is 18.54%. An updated key to world species of Vitronura is also provided.