As well as other animals, honeybees are often infected by microorganism included sacbrood virus (SBV). The early infective SBV detection and diagnosis became necessary for control of SBV. In this research, the Realtime-PCR (qPCR) technique was applied in comparison with RT-PCR technique in which the total RNA were extracted by TRlzol-LS. The results of examination of 18 honeybee samples that analysed with both techniques were showed with significant differences, in which qPCR was positive detection with 17/18 samples (94.44 percent) while RT-PCR technique was detected only 6/18 positive samples (33.33 percent). In addition, the experiment, qPCR was showed the high capacity for SBV detection even with the dilutation concentration of 10-6 that is more sensirive in comparison with RT-PCR technique.