This study was carried out to investigate the prevalence of honeybee (Apis mellifera) disease in cheonan and asan area. From September to November in 2012, 33 samples were collected from 33 apiculture farms in the regions and reverse transcriptase-polymerase chain reaction (RT-PCR) and polymerase chain reaction (PCR) was conducted. Among 33 samples, prevalence rate was 42% in Sac Brood Virus (SBV), 52% in Nosema, 21% in American foulbrood (AFB), 70% in European foulbrood (EFB), 97% in Stonebrood, 3% in Chalkbrood. The result indicate that stonebrood was most prevalent disease in apiculture farms in cheonan and asan area.