| 摘要: |
| 为建立非洲马瘟病毒(African Horse Sickness Virus,AHSV)的血清学诊断方法,本研究构建了非洲马瘟病毒VP7蛋白的原核表达质粒pET-32a-VP7,诱导表达后使用Ni-NTA对可溶性VP7蛋白亲和纯化,以重组VP7蛋白作为包被抗原,优化建立检测血清抗体的间接ELISA方法,并对该方法进行敏感性、特异型和重复性验证,表明该方法的检测效果良好。本研究为非洲马瘟的血清检测试剂盒的开发提供的相关参考。 |
| 关键词: AHSV VP7 酶联免疫吸附试验 检测 |
| DOI： |
| 投稿时间：2023-07-06修订日期：2023-11-08 |
| 基金项目:“十四五”国家重点研发计划2022YFD1800500；中国兽医药品监察所兽药行业公益性重点专项“非洲马瘟病毒毒种研究”(GY202030) |
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| Prokaryotic expression of African horse sickness virus VP7 protein and establishment of an indirect ELISA antibody detection method |
|
| (China Institute of Veterinary Drug Control) |
| Abstract: |
| In order to establish a serological diagnosis method for African Horse Sickness Virus (AHSV), a prokaryotic expression plasmid pET-32a-VP7 was constructed. After induction of expression, soluble VP7 protein was affinity purified by Ni-NTA. The indirect ELISA method was optimized and established using the recombinant VP7 protein as the coating antigen. The sensitivity, specificity and repeatability of the method were verified, and the detection effect of the method was good. This study provides a reference for the development of serum detection kit for African horse sickness. |
| Key words: AHSV VP7, Enzyme linked immunosorbent assay Detection |
