We used a non-invasive technique with microsatellite primers to investigate genetic variation among Eurasian otters Lutra lutra in eastern South Korea. We collected twenty two otter spraints in January and six in August 2008. We used spraints from five dead otters from five different river systems for the present genetic analysis. We extracted DNA from 20 spraints from the January sample. Ten microsatellite primers (Lut435, Lut453, Lut457, Lut604, Lut615, Lut701, Lut715, Lut717, Lut733, and Lut832) for Eurasian otters were tested, and four loci were successfully amplified for further analyses. The results of genotyping the otter population with microsatellite loci lead to the identification of 9 individuals from the Ungokcheon Stream. The Ungokcheon population also showed a genetic structure represented by the Hardy-Weinberg equilibrium.