Pathogenicity and Molecular Characterization of A New Infect

发布时间：2023-02-15 19:44

　　

【文章页数】：138 页

【学位级别】：博士

【文章目录】：
ABSTRACT List of abbreviations CHAPTER 1 Review of Infectious Bronchitis Virus and Infectious Bronchitis Disease
1 Introduction
    1.1 Avian Infectious Bronchitis Virus (IBV)
    1.2 Classification
    1.3 Structure and morphology
    1.4 Genomic organization
    1.5 Non structural and structural proteins
    1.6 Tissue tropism
    1.7 Virus entry and attachment
    1.8 Replication, transcription and translation
    1.9 Virus assembly and release of particles
    1.10 Prevalence and propagation of IBV
2 Infectious Bronchitis Disease (IB)
    2.1 IBV serotype
    2.2 IBV genotype
    2.3 IBV protectotype
    2.4 Susceptible Host
    2.5 IBV intra-classification
    2.6 Identification of Virus
    2.7 Pathogenicity
    2.8 Morbidity and mortality
    2.9 Clinical signs
    2.10 Necropsy changes
    2.11 IBV vaccines and immunity
3 IBV diagnosis
4 Aims and objectives of the study
References CHAPTER 2 Epidemiology of infectious bronchitis virus (IBV) and control measures
1 Introduction
    1.1 The origin of IBV
    1.2 Worldwide epidemiology of infectious bronchitis virus
    1.3 IBV epidemiology in China
2 IBV QX strain emerging worldwide
3 IBV control measures
    3.1 IBV back ground knowledge
    3.2 IBV control strategies
References CHAPTER 3 Pathogenicity of an infectious bronchitis virus isolate in China
1 Introduction
2 Materials and methods
    2.1 Collection of virus samples
    2.2 Specific pathogen free embryonated eggs and chicken
    2.3 Identification of IBV by RT-PCR
    2.4 Determination of 50% embryo infectious dose (EID₅₀)
    2.5 IBV experimental infections to chicken
    2.6 Histopathological examinations
3 Results
    3.1 Identification of IBV by RT-PCR
    3.2 Determination of 50 percent chicken embryo infectious dose (EID₅₀)
    3.3 IBV experimental infection
    3.4 Morbidity and mortality rate
    3.5 Clinical signs
    3.6 Gross pathological lesions
    3.7 Histopathological lesions
4 Discussion
References CHAPTER 4 Full length genome sequencing of an infectious bronchitis virus isolate XDC-2
1 Introduction
2 Materials and methods
    2.1 Primer designing
    2.2 Total viral RNA extraction,RT-PCR and PCR
    2.3 Race PCR for amplification of IBV RNA genome 5'end
    2.4 IBV fragments amplification, cloning and sequencing
    2.5 Separations of structural and nonstructural genes
    2.6 IBV XDC-2strain full length genome pairwise comparison
    2.7 Phylogenetic tree analysis
    2.8 Comparisons of S glycoprotein gene amino acids with vaccine strains
    2.9 Selection pressure
    2.10 Recombination analyses
3 Results
    3.1 IBV nucleotide sequencing
    3.2 Full length genome nucleotide homology and divergence
    3.3 IBV XDC-2 strain full length genome pairwise comparison
    3.4 Phylogenetic analyses
    3.5 Comparison of S gene amino acids with vaccine strains
    3.6 Analysis of selection pressure
    3.7 Recombination analyses
4 Discussion
References CHAPTER 5 Construction and expression of recombinant plasmids containing IBV S1 and Ii-key segment of chicken major histocompatibility complex Ⅱ gene
1 Introduction
2 Materials and methods
    2.1 Cells
    2.2 Primer designing
    2.3 RNA extraction,RT-PCR and PCR
    2.4 Cloning of the complete IBV S1 gene and chicken MHC Ⅱ Ii gene
    2.5 Preparation of DH5 α E. coli competent cell for transformation of plasmids
    2.6 Plasmid extraction by DNA plasmid purification kit
    2.7 Confirmation of recombinant plasmids
    2.8 Eukaryotic expression of DNA recombinant plasmids
    2.9 SDS-PAGE AND Western blot protocols
    2.10 Chicken, plasmids and vaccination
    2.11 Detection of anti-IBV specific antibodies
    2.12 Li gene specific T lymphocyte cells proliferation assay
    2.13 Statistics
3 Results
    3.1 Amplification and cloning of IBV S1 gene and chicken MHC Ⅱ Ii gene
    3.2 Confirmation of the recombinant plasmid (pcDNA-S1-F) by colony PCR and Enzyme Digestion
    3.3 Confirmation of the recombinant plasmid (pcDNA-Ii-F) by colony PCR and
    3.4 Confirmation of the expression of the recombinant plasmids by Western blot analysis
    3.5 Induction of immune responses
4 Discussion
Acknowledgements
References Conclusions Suggestions Publications Acknowledgements



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