Isolation And Identification Of Sika Deer's Mucosal Disease Virus

In this study, an unknown virus designated MDVCC-deer6 was isolated from livers of a aborted fetus using MDBK cell line. The virus could multifly on the MDBK cells,and induce regular cytopathic effect (CPE).Observed by negative staining electron microscope, the virus could be seen with typical MDV virions. In biological assay, the hemagglutination reaction test proved that this virus had no any reaction to erythrocyte of chook,rabbit, pig and sheep. In the test of virulence determination of isolated MDVCC-deer6, the result of TCID50 is 10-5.5. Physicochemical assays showed that this viurs was sensitive to choroform and ether. In trpsin tolerance assay,this virus could resist to 1% trpsis at 37C in an hour and multifly ,but the average infection titre of the virus decreased 3.2 titre. In acid tolerance assay,this virus was sensitive to pH3.0 at 37C in 1 hour, and the average infection titre of the virus decreased 4.4 titre. In alkali tolerance assay , this virus was resistant to pH9.0 at 37C in 1 hour, and the average infection titre of the virus decreased little.In heat assay, at 50C,the virus was processed from 30 minutes to 70 minutes and at each condition the viral virulence reduced to some certain degree.Among these conditions,when at 50C in 30 minutes,the average infection titre of this virus decreased 0.6 titre,and when at 50C in 70 minutes, the average infection titre of this virus decreased 3.7 titre.at 56C,the virus was processed from 30 minutes to 70 minutes and at each condition the viral virulence reduced to some certain degree too. Among these conditions, when at 56C in 30 minutes,the average infection titre of this virus decreased 2 titre, and when at 56C in 70 minutes ,CPE of this virus disappeared and the virus lost the multiplication capacity completely. By neutrolizaion assay,MDVCC-deer6 could be identified as a kind of MDVBy RT-PCR method,we selected a pair of primers named p1 and p2 which located in highly conserved P125 nucleotide region . we could get 402 bp nucleotide fragement in C24V and 672 bp nucleotide fragement in NADL after amplifying by the pair of primers, we amplified the target fragement of about 400bp from the passages of culture of this isolated virus and the the passages of culture of this C24V. However, the control cell showed negative result.From the results described above, it can be concluded that the strain MDVCC-deer6 is a new strain of deer-origined MDV.