Studies on Molecular Biology, Monoclonal Antibody and DNA Vaccine of Newcastle Disease Virus

• Main Authors: Xin-wei You, 游信偉
• Other Authors: Maw-Yeong Lin
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/86116403678473765062

碩士 === 國立屏東科技大學 === 獸醫學系所 === 95 === Newcastle disease (ND) is a highly contagious viral disease in causing respiratory, digestive and nervous disturbance in domestic poultrys and birds. Phylogenetic analysis of the 13 isolates ND virus (NDV) of 1980s revealed that 9 and 4 isolates possess the sequence of 112RRQKRF117 and 112GK/RQGRL117 respectively at the cleavage site of the F glycoprotein and could be grouped into pathotype of viscerotropic velogenic NDV (VVNDV) (subgenotype Ⅱ, Ⅵf, Ⅵg and VIIe) and pathotype of mesogenic or lentogenic NDV (sugenotypes Ⅰand Ⅱ). The divergence of nucleotide sequence among the isolates of each individual genotypesⅠ, Ⅱ, Ⅵf, Ⅵg type NDV is 0.0%, 0.0-3.5%, 0.6-5.8%, 0.6%. Twenty four isolates of NDVs were antigenically characterized into A, B, C and D groups by using 14 monoclonal antibodies (mAb) prepared from TW/99-V157 isolates. Among them, seventeen isolates reacted with all the mAb were classified into group A. Two isolates neglectly reacted with site 3（157F/9）mAb were classified into group B. Four isolates neglectly reacted with site 1（157F/4 and 157F/5）, site 2（157/3）and site 3（157F/9）mAb were classified into group C, One isolate neglectly reacted with site 2（157F/7and 157/1）mAb were classified into group D. F and HN glycoprotein genes of NDV TW/02-302 were cloned into a mammalian expression plasmid vector (pcDNA 3.1) for expression of HN and F glycoprotein in the transfected vero cell. The above 2 proteins localized on the membrane surface of vero cells can be detected by the by the mAb（157F/2） against TW/02 -V302 NDV-F protein and the binding capability with chicken RBC at 4 days post transfection. Although low level of serum neutralization test (SN) antibody can be detected in the chickens immunizized with variant concentrations of pcDNA3.1/V5-His/TW/02- V302 HN and pcDNA3.1/V5-His/TW/02-V302 F at 3 or 4 weeks post immunizatioin, no any protectivity against the NDV challenge was found in the chickens.