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Establishment of Detection Methods for Hog Cholera Virus with SPA-COA and ELISA

Author: QinTong
Tutor: YangRunDe
School: Agricultural University of Hebei
Course: Preventive Veterinary Medicine
Keywords: Classical swine fever virus Immunoelectrophoresis SPA coagglutination test Double sandwich enzyme-linked immunosorbent assay Detect
CLC: S854.43
Type: Master's thesis
Year: 2006
Downloads: 175
Quote: 2
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Abstract


In this study, PK-15 cell proliferation the the swine fever Shimen Department of virulent F 114 strains (CSFV-F 114 ), through the preparation of calf testis cell proliferation swine fever China lapinized The attenuated C strains (lapinized Philippines Coronel), HCLV (). After repeated freezing and thawing of the virus liquid was HCLV strains using polyethylene glycol 6000 was concentrated and purified obtain high purity HCLV. Concentrated purified virus solution were used to immunize rabbits and pigs the of rabbit anti HCLV and pig anti HCLV hyperimmune serum was prepared by saturated ammonium sulfate precipitation, dialyzed, of of rabbit anti HCLV and pig anti HCLV immunoglobulin IgG in a crude extract. A protein-rich gold standard strains of Staphylococcus aureus (Cowan I) based on the characteristics of the Staphylococcus aureus cell wall protein A (SPA) labeled rabbit anti HCLV and the pig anti HCLV hyperimmune serum, to establish the diagnosis of classical swine fever SPA tablet coagglutination test. SPA diagnostic reagents with different dilutions of the anti-serum preparation, co-agglutination test the original times serum, rabbit anti HCLV, swine anti HCLV positive serum 2-fold dilution resulting diagnostic reagents best. Compare diagnostic sensitivity of the solution of two serum markers SPA, the diagnostic fluid antigen HCLV SPA Zuigao Jian out rabbit anti-concentration: 1:32, the pig anti the diagnostic reagents antigen of HCLV SPA Zuigao Jian out concentration: 1:64. And convection immunoelectrophoresis to 1:2 times diluted antigen is detected compared to the higher sensitivity of the method. By the detection of the sensitivity of the incidence of animal organs, the proof of the lymph nodes and spleen in the diagnosis of classical swine fever in the clinical use of SPA coagglutination assay for the best detection site. In addition, the operation is simple, fast, save material, without special equipment, new technology is a suitable for rapid diagnosis and epidemiological investigation of the grassroots pig swine fever. Using rabbit anti-the HCLV IgG, swine anti-HCLV IgG and goat anti-rabbit enzyme-labeled antibody, the establishment of the double sandwich ELISA for detecting CSFV antigen immunological methods. The experimentally determined rabbit anti lapinized Philippines Coronel), HCLV (IgG, swine anti HCLV IgG and the best use of the enzyme-labeled antibody concentrations were 1:40,1:200,1:400. The sensitivity of the test, a specific inhibitor of crossover trial repeatability test results showed that this method has high sensitivity, specificity, reproducible advantages. Is a suitable laboratory confirmed swine fever virus. The application of this study to establish the two methods of detecting CSFV antigen detection of 50 cases of clinical samples. SPA coagglutination test detected positive samples were positive rate of 12%, results can be observed to 7min within,; sandwich ELISA detected positive samples 10 were positive rate of 20%, the time required is approximately 24h .

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Veterinary clinical medicine > Veterinary diagnostics > Laboratory diagnostic method
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