Amplified fragment length polymorphism (AFLP) technology was used to evaluate the genetic diversity of three introduced populations (Yantai, Dalian and Laizhou) of introduced barfin flounder(Verasper moseri). The genetic distances and identity index were calculated, and according to them, the UPGMA dendrogram was constructed. A total of 827 loci ranged from 50－700 bp were detected using 10 primer combinations, and 8－37 polymorphic loci were detected per primer combination. The percentage of polymorphic loci of three introduced populations of barfin flounder were 29.14%, 15.60% and 20.31%, respectively. The Shannon’s information index were 0.1799, 0.0949 and 0.1231, respectively, and the Nei’s gene diversity were 0.1225, 0.0658 and 0.0848, respectively. The total genetic diversity level of three introduced populations was low, of which YT population was the highest, LZ population was lower, and DL population was the lowest. The genetic distance between YT introduced population and DL introduced population was the highest(0.0230), whereas, the genetic distance between LZ introduced population and DL population was the lowest(0.0129). The coefficient of gene differentiation(Gst) value was 0.219, showing a certain extent of differentiation among three populations of Barfin flounder.