發展檢測犬小病毒檢測試劑

• Main Author: 蔡欣憶
• Other Authors: 莊國賓
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/24531779250091088907

碩士 === 國立屏東科技大學 === 動物疫苗科技研究所 === 100 === Canine parvovirus (CPV) is a small and non-enveloped icosohedral symmetry virus, containing a 5.2kb single-stranded DNA. The virus is from feline panleukopaenia virus (FPLV) throµgh a small number of mutations in the single capsid protein. There are two serotypes of Canine Parvovirus. The type 2 of canine parvovirus (CPV-2) causes hemorrhagic enteritis and sometimes myocarditis in dogs. Dogs are infected through oral contact with CPV2 in feces, infected soil, or fomites that carry the virus, following by ingestion of the feces, hence, the virus replicates in the lymphoid tissue in the throat, and then spread to the bloodstream. From there, the virus attacks rapidly dividing cells, notably those in the lymph nodes, intestinal crypts, and within bone marrow. We isolated the CPV from infected dogs and amplified the VP2 fragments by PCR. And then, the VP2 gene of CPV was constructed into the E. coil expression vector and purified by Ni2+ affinity column. The antigenicity of recombinant VP2 protein was characterized by Western blot with Anti-HIS antibody and CPV infected dog serum. The Anti-CPV-VP2 was screened 13 monoclonal antibody and polyclonal antibody . single out of which a number Anti-CPV monoclonal antibody-6 field strains with excellent adhesive force. The Anti-CPV polyclonal antibodies were coated on the ELISA plate and 10 dogs’ faecal specimens were used as the antigene. Then the captured CPV were probed with Anti- CPV monoclonal antibody-6 in sandwhich ELISA. Then the captured CPV were probed with Anti-CPV monoclonal antibody in sandwhich ELISA. The faecal samples were used for CPV-VP2 gene detection by PCR. demonstrates that polyclonal antisera gave visually positive reactions with 12 out of 14samples. we need more wild type sample to test The strip of evaluate the specificity and Sensitivity. The were generated and used for strip development for pre-vaccination screening and disease prevention.