| 摘要: |
| 目前狂犬病疫苗的效力检验采用NIH法，需要使用狂犬病毒强毒株进行攻毒试验，具有一定的生物安全风险。为建立检测小鼠血清抗体效价的间接ELISA方法，代替NIH法中的脑内攻毒试验，扩增狂犬病毒（RV）G蛋白基因，并将其克隆至大肠杆菌pET-32a载体上进行表达。以该蛋白作包被抗原，摸索试验条件，建立检测小鼠血清抗体效价的间接ELISA方法。使用此方法与国际公认的荧光抗体病毒中和试验（FAVN）法比较，两者检测结果曲线相关系数为0.986，表明两者相关性较好，但ELSIA方法更加快捷、简便。本试验建立的间接ELISA方法可用于检测小鼠血清中狂犬病抗体，为狂犬病疫苗效力检验替代方法的建立提供了基础。 |
| 关键词: 狂犬病，G蛋白，原核表达，ELISA，替代方法 |
| DOI： |
| 投稿时间：2022-01-06修订日期：2022-03-03 |
| 基金项目:十三五国家重点研发计划项目(2016YFD0501004) |
|
| Prokaryotic Expression of Rabies G Protein and The Establishment of its Indirect ELISA |
|
| (China Institute of Veterinary Drugs Control) |
| Abstract: |
| The effectiveness of rabies vaccines is tested using the NIH method, which requires the use of virulent strains of rabies virus for challenge tests, which poses a certain biological safety risk. To establish an indirect ELISA method for detecting antibody titers in mouse serum to replace the brain challenge test in the NIH method. The G protein gene of the rabies virus (RV) is amplified and cloned into the E.coli pET-32a vector for expression. Indirect ELISA method for detecting antibody titer in mouse serum has been established with the expressed G protein. Compared with the fluorescent antibody virus neutralization test (FAVN), which is internationally recognized, the correlation coefficient of the two methods is 0.986, demonstrated that these two methods have highly consistency. Comparatively, the ELISA is simpler and more efficient. The indirect ELISA method established in this experiment can be used to detect rabies antibodies in mouse serum, providing a basis for the establishment of an alternative method for testing the efficacy of rabies vaccines. |
| Key words: Rabies, G protein, prokaryotic expression, ELISA, alternative method |
