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Study on Monoclonal Antibody to PRRSV GP5 Protein and DNA Vaccine of apx IA Gene of Actinobacillus Pleuropneuromoniae

Author: XieTianTian
Tutor: ChenHuanChun
School: Huazhong Agricultural University
Course: Biochemistry and Molecular Biology
Keywords: Porcine reproductive and respiratory syndrome virus GP5 protein Monoclonal Antibodies Actinobacillus pleuropneumoniae apx Ⅰ A gene DNA vaccines
CLC: S858.28
Type: Master's thesis
Year: 2005
Downloads: 285
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Abstract

Porcine reproductive and respiratory syndrome (porcine reproductive and respiratory syndrome, PRRS) is a porcine reproductive and respiratory syndrome virus (PRRSV) caused by a new infectious disease, the clinical to sow fever, anorexia, premature birth, miscarriage, mummified, stillbirth, low earners and other reproductive disorders and respiratory symptoms pigs of all ages and is characterized by high mortality rates of piglets. The disease was first reported in 1987 in the southern United States, is now available throughout the world, to the global pig industry caused huge economic losses. Immunization is a major disease prevention and control measures. Safe and efficient vaccine depends on the PRRSV-depth understanding of molecular immunology. Therefore, we must establish a virus \Monoclonal antibody is such marking systems. Since PRRSV and other reproductive disorders (such as swine fever, porcine parvovirus, Japanese encephalitis, swine pseudorabies, etc.) in clinical practice is very similar, and often occurs with other bacterial diseases common infections, while different farms clinical PRRS infection after another big difference. Based solely on clinical symptoms and epidemiological characteristics is difficult to make an accurate diagnosis, the diagnosis also requires the help of laboratory diagnosis. Establish a high sensitivity and specificity, rapid diagnostic methods, monoclonal antibody is a good choice. In this study, the expression of GST fusion porcine reproductive and respiratory syndrome virus GP5 protein as an immunogen, and the establishment of a stable and prepared for the secretion of porcine reproductive and respiratory syndrome virus GP5 protein monoclonal antibody hybridoma 1H11C4, its cell culture supernatant titer of 1:2 ^{9 , mouse ascites titer of 1:100 × 2 ^{11 . ELISA and Westren-blot results showed that, 1H11C4 only fusion expression of GST-GP5 protein response, but with KG protein GST not react. Indirect immunofluorescence showed 1H11C4 with PRRSV responses obtained in this study indicate that monoclonal antibody is specific for PRRSV GP5 protein. Further use of recombinant GP5 protein deletion mutants determined that mAbs mainly for GP5 protein C-terminal 67 ~ 200 aa. Epitopes on and found that monoclonal antibodies can enhance PRRSV in MARC-145 cells proliferation titers This is for further study of PRRSV GP5 protein of molecular biology and immunological characteristics laid a solid foundation. Actinobacillus pleuropneumoniae (Actinobacillus pleuropneuromoniae, APP) pleuropneumoniae caused serious harm to the pig industry is a respiratory infection, and its extensive fibrous acute cases of pulmonary hemorrhage is characterized by chronic lung focal necrosis and pleurisy is characterized with high morbidity and mortality. The disease was distributed worldwide to the pig industry has caused tremendous economic losses. Vaccine is considered to be the best choice for disease prevention and control, in order to prepare the country isolates the local epidemic strains of inactivated vaccine infection can provide better protection force, this vaccine serotypes for the same kind of infection can reduce mortality, but it can not reduce morbidity, and chronic lung disease can not prevent infection, but can not provide cross-protection against atypical bacteria, therefore, an urgent need to develop new vaccines and diagnostic methods to prevent and treat the disease.}}

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