Rabbit Haemorrhagic Disease Virus Partial Gene Sequencing And Experimental Infections Of DJRK

A PCR -ELISA was established to detected Rabbit Hmorrhagic Diease Virus (RHDV) in infected DJRK cell ,A pair of primers were sesigned to amplify a fragment of the gene encoding the structural polypeptide VP60.A probe matched the interior of the amplified fragment was modified with biotin at its 5"-end.The best conditions of PCR-ELISA were:appropriate cycle mumber of amplification was 25;hybridization temperature and time was 40℃ and 3h respectively.The sensitivity of detection of DIG-PCR products is two times higher than electrophoresis in agarose gel stained with ethdium bromide .The cut-off limit tested of a postive result is A405^0.200.The result RHDV could infect DJRK and generate was testified.Virus genome started to duplicate at 8h post-infection and then to augment rapidly and to arrive a most amount at 20h post-infection. But HA couldn"t be inspected and CPE couldn"t be observed.RHDV could propagated on the cell cycle stably Seven Rabbit Haemorrhagic Diseas Virus(RHDV) strains were obtained from different area or period in china.Two oligonucleotide primers were designed to amplify interior of the gene encoding the structural polypeptide VP60.PCR products were sequenced. Multiple sequence alignments were carried out with ClustalW.The result revealed o high degree of homology between seven of the isolates and the reference strain FRG. Despite the high homology,variation was still demonstrated and appeared to be related to the times.Amino acid alterations translation from RNA also sustain the conclusion.