| Duck virus enteritis(Duck Virus Enteritis, DVE), also known as duck plague(Duck Plague, DP), is caused by duck virus enteritis(Duck Enteritis Virus, DEV). DEV is an acute, heat and septicaemic contagious viral disease which can infect ducks, geese and a variety of Anseriformes birds. DEV was classified as a thired-class epidemic by Chinese Law on Animal Epidemic Prevention, and it was one of the main diseases against the current development of poultry industry, characterized by the prevalence of extensive and rapid spread, high morbidity and mortality. Duck virus enteritis is unclassified viruses belonging to Herpesviridae .The diameter is 160~180 nm, and genome size is about 150 Kb, and covalently connected by a long unique areas(UL) and unique short zone(US). Glycoprotein B is coded by ul27. The length of ORF is 3003 bp, containing 1000 amino acid residues, and molecular weight is 113.7 KDa. In all of the herpes simplex virus, gB is an essential glycoprotein in virus adsorption, penetration and cell-mediated integration. While gB is the major target antigen in the host immune response, can induce cellular immunity and stimulate the body producing neutralizing antibody and cytotoxic T lymphocyte response. The function of duck enteritis glycoprotein B is not knowed, then, preparation of the monoclonal antibodies of glycoprotein B of duck enteritis virus is not reported.Construction of the duck enteritis virus glycoprotein B(DEV-gB) eukaryotic expression vector: pcDNA3.1-DEV-ul27 as antigen for immunization, united with bupivacaine hydrochloride immuned BALB/c mice, and seted up indirect immunofluorescence method for screening of anti-DEV glycoprotein B monoclonal antibody. Spleen cells were seperated ,and fused with SP2/0 myeloma cell, using the equally distributing cells method, after cloning 3~4 times, finally won a total of 13 secretion of monoclonal antibody(McAb) named the hybridoma 2G7, 3D7, 5G5, 6A6, 6D1, 6E10, 8A2, 9B8, 9F4, 10H2, 11A5, 11B4 and 12G9. Chromosome number was 92~110 strips, the secretion of monoclonal antibodies subclasses were IgM, light chain wereκ. The monoclonal antibodies can be long-term culturee in vitro, and can secrete sustained and stable antibody .The monoclonal antibodies in this experiment as the first antibody, with 9 truncated fusion proteins which had antigenic sites predicted by biology software, Western blot analysis was done to analyse DEV-gB epitopes.The result were that, 2G7, 6A6, 6D1, 6E10, 8A2, 9F4 and 11B4 all can specific identify 61~110 aa; and the monoclonal antibody 3D7, 5G5, 11A5, and 12G9 epitope recognition region was located within 655~720 aa. This 9B8 and 10H2 monoclonal antibody do not respond to truncated proteins, may be targeted epitopes are conformational.The reactivity between monoclonal antibodies and the whole virus particles were detected by Dot-ELISA. 8 strains monoclonal antibody were testified reaction with the natural virus, which were: 3D7, 5G5, 6E10, 6D1, 8A2, 9B8, 10H2 and 11B4.The result indicated that 3D7, 6D1, 8A2, 9B8, 11A5 and 12G9 had neutrality activity through microdosis cell affection neutrality experimentation and lacunae reduction neutrality experimentation to identify neutrality activity of gained monoclonal antibody. The titer of neutrality activity of 3D7 and 6D1 were 100, others were 10.Preparation of the monoclonal antibodies of glycoprotein B of duck enteritis virus, and identified to Western blot analyze, its reaction with virus and neutralization activity. Some monoclonal antibody can be used to analyze glycoprotein B antigen epitope with further brachytmema expression glycoprotein B; the function of natural glycoprotein B peptide and antigen epitope will be identified using the reaction of theas monoclonal antibody with brachytmema expression protein; monoclonal antibody which is material foundation can be used to establish detected method, for example: Indirect Immuno-Fluorescent and enzyme-linked immunosorbent assay(ELISA) to detect antibody of illnes duck.
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