Metcalfa pruinosa (Say), native to North America, is spreading rapidly in the Korean Peninsula, causing serious damages on many deciduous forest trees, ornamental trees, and agricultural crops. Before the first report from Korea in 2005, M. pruinosa has not been reported from any other countries in the Eastern Palaearctic, while it has been record in Italy in 1979, and rapidly spreading into many European countries. To trace the invasion route of this species, we analyzed haplotype analysis of the mitochondrial cytochrome c oxidase subunit-I (mtCOI, 577bp), and developed microsatellite markers. In the haplotype analysis, a total of fourteen haplotypes were found from 69 individuals in 18 populations: 12 haplotypes (native region), 4 haplotypes (European region) and 2 haplotypes (Korea). Interestingly, Korean populations were clustered with some European populations. Eight polymorphic microsatellite loci were developed, and population structures were analyzed from 145 individuals in 8 populations. The origin and invasion route of M. pruinosa are under investigation.