Abstract: The RT-PCR assay for detection of Rabbit hemorrhagic disease virus (RHDV) in nasal swab specimens collected from rabbits was developed for the research of virus carrying in healthy immunized rabbit. A pair of primers were designed according to the conserved sequences of vp60 from RHDV published in GenBank, and a 591 bp segment was amplified by RT-PCR. The least amount of RHDV could be detected was 2.40 ng·μL^-1, and the sensitivity of the RT-PCR was 8×103 times higher than that of HA method. 168 nasal swabs collected from 5 provinces was detected by the assay, the results showed that the positive samples was 22 and the positive rate was 13.09%. These results indicated that the phenomenon of virus carrying in healthy immunized rabbit and the developed RT-PCR might be a promising approach in clinical screening, detection and carrying research of RHDV in a large number of immunized rabbit nasal swabs.