We characterized 16 novel polymorphic loci isolated from a partial genomic DNA library of Odontobutis potamophila enriched for CA repeats. We tested the variability of these microsatellites on 51 unrelated individuals collected in China. All loci were polymorphic. The average allele number was 14.6 per locus, ranging from 2 to 27. The observed heterozygosity ranged from 0.35 to 0.90, with an average of 0.70, whereas the average expected heterozygosity was 0.76. Twelve of the 16 microsatellites conformed to Hardy-Weinberg equilibrium and were inherited independently. These developed microsatellites will be useful in studies of population genetics and other genetic studies on this important food species.