Canine Parvovirus:Isolation,Identification,Developing Its Monoclonal Antibody And Application

The faeces from dogs detected with haemorrhagic enteritis by haemagglutionation assay (HA) and confirmed inhibition test (HI) were positive. Infected small intestine was determined by detection of intranuclear inclusion bodies in cells .The embryonic feline kidney cell line (FKS I) was used to isolated viruses. The cell supernatant fluid was detected b HAIHI. Canine parvo~抜rus particles were detected by electron microscope (EM). Specific fluorescence was apparent only in infected cells by immunofluorescence assay(IFA) detected. The above results indicated that CPV was isolated from dog with haemorrhagic enteritis and was identified. CPV-YZ strain was screen.it resisted acid(PH3). trypsin. and chloroform but was sensitive to formaldehyde and heat(80C), its TCID5O was O.lml ~ SDS-PAGE revealed that the virus contained 2 structure polypeptides: VPI(76KD).VP2(64KD). with VP2 as the major structural protein. Eight hybridoma cell lines secreting monoclonal antibody(McAb) against CPV have been established. They were respectively named F5(; 10. 3F 11 Fl . F 11 (;6. 6(75(22. 6H(;3. 6(221311 . Fl I 1)6~ 31)9(71. McAbs subclass is lgG. All were analyzed b Dot-Ei.ISA. McAb only binds to viruses that have three-dimension structure. In comparison. LA has lower sensitive and easier to pertorm than HA. The infected cell was detected in the same time b~?HA and IFA.In the immuoenzyme histochemistrv test .the monoclonal antibody could react with the viruses and those brown dots in the tissues of the organs could be seen.