Origin of the red imported fire ant (RIFA, Solenopsis invicta) is South America, a tropical climate region. It has settled in the invasive area, causing various problems. In recent, colonies of Solenopsis invicta were discovered in Busan in 2017, and then in Busan, Pyongtaek, Incheon, Daegu and Ansan in 2018. This study aims to validate 31 microsatlite loci referred from previous study for tracing origin of Solenopsis invicta colonies. We tried to test previously developed microsatellite loci so that we establish the protocol for molecular epidemiology of RIFA. We applied 31 microsatellite loci to DNA samples from the RIFA colonies found in Texas and Florida, USA. We will present 31 markers were divided into three injections and the amplification rate for each marker was shown as follows.