Development of an Indirect ELISA with Artificially Synthesized N Protein of PPR Virus
The full-length gene encoding the nucleocapsid (N) protein of the virus (PPRV) responsible for an outbreak of peste des petits ruminants in Tibet in 2007 was synthesized in two stages using overlapping PCR without the need for viral genomic cDNA as template. The full-length N gene was successfully expressed in Escherichia coli， and the purified gene product bound to monoclonal antibody raised against PPRV N protein. Furthermore, it was able to replace recombinant B-N antigen as the coating antigen in a commercial ELISA kit prepared with another PPRV strain. Recombinant protein was employed as the coating antigen to develop an indirect ELISA for PPRV antibody detection in the sera of infected small ruminants. Antibody detection was optimal at a 1：200 serum dilution and an antigen concentration of 3.2 g/ml, and the positive threshold (cutoff) value of the assay was 2.18. Analysis of 697 serum samples revealed the sensitivity and specificity of the indirect ELISA to be 96.7 and 96.1％, respectively, compared with a commercially available ELISA test.
小反刍兽疫病毒 核衣壳蛋白 人工基因合成 抗体检测 酶联免疫吸附试验
Guo-rui Zhang Jiang-yong Zeng Yu-min Zhu Shi-juan Dong Se Zhu Rui-song Yu Ciren Duoji Zhi-hai Lei Zhen Li
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing Tibet Livestock Research Institute, Tibet Academy of Agricultural and Animal Sciences, Lhasa , China Institute of Animal Sciences and Veterinary Medicine and Shanghai Key Laboratory of Agricultural Gen
国内会议
上海
英文
1-9
2011-10-01（万方平台首次上网日期，不代表论文的发表时间）