| Summary: | 碩士 === 國立屏東科技大學 === 獸醫學系所 === 96 === Canine parvovirus type2 (CPV-2) heamorrhagic enteritis has been an important infectious disease in canine since its first outbreak in 1978 in USA. The disease has been found occurred in all ages of canine especially in puppy in causing myocarditis, leukopenia, vomiting, heamorrhage diarrhea and even death. Canine parvovirus (CPV) is a member of the Parvoviridae. The genome of CPV is a single stranded DNA of negative-sense with 5.2 kb in size. The genome of CPV encodes VP1, VP2 and VP3 structural proteins, and NS1, NS2 nonstructural proteins. The VP2 glycoprotein is the main capsid protein with 6 to 8 antigenic epitopes in inducing the neutralization antibody to the host.
The full length of CPV-VP2 gene (1755 bp) in 10 Taiwan local isolates of 2004 to 2006 and 3 commercial vaccine strains were amplified by polymerase chain reaction (PCR), cloned and sequenced for phylogenetic analysis of their nucleotides and amino acids by DNAStar and Mega 3.1 with the other CPV isolates deposited in the Genebank. Divergeuce on the sequence of amino acid between 10 local isolates and 3 vaccines strains of CPV ranged from 1.7％ to 3％. The 10 local isolates were sufgrouped as CPV-2b based on 2 changes on the VP2 protein at 297(S→A) and 300 (A→G) this might be the main reason in causing the vaccination fail in Taiwan.
The full length of the CPV-VP2 subunit protein of CPV-306/TW05 isolate was successfully expressed in Pichia pastoris. The recombinant subunit protein with molecular weight of 72.7 kDa was obtained and Western blotting characterized. This subunit protein was further mass produced as antigen for subunit vaccine evaluation in beagle puppies.
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