Acute bee paralysis virus (ABPV) is a small single stranded RNA virus recently classified within the family Dicistroviridae, genus Cripavirus. Here, we describe the first study of ABPV in unhealthy bee colonies, which has been an unusual loss in adult bee population and significant honey bee mortality during the year. The aim of this study was evaluation of ABPV infection in honey bee colonies in apiaries with loss of population in different geographical provinces in Iran. Adult bee samples were collected between July - September 2011 and 2012 and were originated from 23 provinces with different geographic areas of Iran. Following the RT-PCR reaction with the specific primers on the isolated RNA, an approximately 618 bp product was detected. We demonstrated the presence of ABPV RNA in 9 (5.62 %) out of 160 samples collected from Iranian apiaries.