Alternative Title鶏アデノウイルス病原性の遺伝子解析および診断用抗原と抗体の開発
Note (General)Fowl Aviadenovirus (FAdV) belong to the family Adenoviridae and the genus Aviadenovirus. FAdV is a non-enveloped icosahedral viros which has a linear, double- stranded DNA. It has been grouped into five species and 12 serotypes. FAdVs infections often cause Hydropericardium syndrome (HPS), inclusion body hepatitis (IBH), egg drop syndrome (EDS), gizzard erosion (GE), and respiratory tract disease. The disease occurs worldwide leading to great economic losses in the poultry industry. To elucidate factors associated with the virulence of Fowl Aviadenovirus, whole genome sequencing using Next Geneneration Sequencing (NGS) of FAdV A strain JM1/1 was carried out. Developing the non-structural protein based Fluorescent antibody virus neutralization (FAVN) and Western Blotting assay (WB) coupled with Immunofluorescence assay (IFA) were studied. The results shown as following 1) The complete genome of FAdV A strain JM1/1 is 43,809 nucleotide length. It’s revealed 99% nucleotide sequence identical to FAdV A strain CELO (chicken embryo lethal orphan), which is an European apathogenic reference strain. The nucleotide sequence differences were analyzed, interestingly showing multiple sites insertions and deletions. The results will provide information on the evolution and may help elucidate viral pathogenesis on molecular biology especially on genetic roles. 2) The recombinant DBP was constructed and subjected as the primary antibody in the developed FAVN test. A DNA Binding Protein (DBP), a non-structural protein, which is detectable early and responsible for initiating DNA replication. DBP was found to be a more conserved domain region within the FAdV serotype 1. The developed FAVN test was compared with a conventional VN test by examining the antibody titer in field chicken sera. The results showed the measured neutralizing antibody titers were high correlated with the VN as the correlation coefficient was 0.8. This FAVN test is simple, achieved quickly, easily and could be an alternative test for FAdV infection. 3) Likewise, the recombinant 52K was constructed and subjected to use as primary antibody in the developed test based on WB and IFA. 52K, non-structural protein, involve in capsid assembly and/or genome packaging. It is especially expressed in late stages of viral life cycle. The C terminal region of 52K was found to be a more conserved domain within all serotypes. WB and IFA analyze revealed that anti-52K antibody can detect FAdVs infection including homologous and heterologous serotypes. Therefore, this antibody possesses a property which could play a role in an alternative method for FAdVs infection diagnosis. In conclusion, this research study provided some knowledge, which will be useful in investigation, surveillance, and elimination of FAdV diseases.
Collection (particular)国立国会図書館デジタルコレクション > デジタル化資料 > 博士論文
Date Accepted (W3CDTF)2019-05-06T10:27:56+09:00
Data Provider (Database)国立国会図書館 : 国立国会図書館デジタルコレクション