Honey bees are critical for agricultural crops pollination. However, many countries have reported high annual colony losses caused by multiple possible factors. Diseases, including those caused by viruses of honey bees, are a significant cause of colony losses. However, nothing is known about the occurrence of viruses of honey bees in Palestinian honey bees. Therefore, eighty colonies from eight apiaries in Bethlehem and Hebron governorates were randomly selected and screened for the presence of 15 viruses of honey bees via quantitative PCR technique. Ten viruses were detected among which the Black Queen Cell virus (BQCV) was the most prevalent (96 % of all colonies) followed by Deformed Wing viruses (DWV) A (77%) and B (72%) and a new emerging virus, the Big Sioux River virus (BSRV, 70%). Other viruses (Apis mellifera filamentous virus (AmFV), Lake Sinai virus 1 (LSV-1), Acute Bee Paralysis virus (ABPV), Sacbrood virus (SBV), Israeli Acute Paralysis virus (IAPV), and Chronic Bee Paralysis virus (CBPV) had lower colony prevalences ranging between 9 -29%. The study showed that most of the viruses were present in all apiaries, and multiple virus infections per colony were common in all colonies. Our findings fill a knowledge gap about the prevalence of viruses of honey bees in Palestine, which could assist in protecting the Palestinian beekeeping industry.