Studies on the use of immunocapture-RT-PCR assay for detection of Polish strains of the European brown hare syndrome virus (EBHSV)

Seven strains of EBHSV and 29 samples of internal organs of dead hares were used in the investigations. A Polish strain of rabbit haemorrhagic disease virus (RHDV) as well as a negative antigen control was sued for comparison. Among 29 samples from dead hares, 3 originated from hares experimentally infected with a standard Italian strain and 1 from a hare infected with the Polish strain, isolated in 1992. Studies confirmed the presence of viruses in 9 out of 38 samples, positive in ELISA and classic method of RT-PCR. Moreover, the presence of EBHS virus was detected by nested PCR conducted with a product of immunocapture-RT-PCR assay (IC-RT-PCR) in a sample originating from a hare infected with Italian strain, although ELISA was negative. IC-RT-PCR assay sensitivity was from 50 to 128-fold higher in comparison to ELISA. The prepared reagents used in IC-RT-PCR assay facilitated the detection of EBHS virus.