Grassi M, Wandeler A I, Peterhans E
Institute of Veterinary Virology, University of Bern, Switzerland.
J Clin Microbiol. 1989 May;27(5):899-902. doi: 10.1128/jcm.27.5.899-902.1989.
The envelope glycoprotein G of rabies virus induces neutralizing antibodies, which are important in protection against rabies. This protein was solubilized from purified virus and isolated by differential and sucrose density gradient centrifugation followed by high-performance liquid chromatography. Conditions for solubilization and purification of G were optimized by using immunoblotting and enzyme-linked immunosorbent assay techniques. The reaction with conventional antisera and monoclonal antibodies indicated that purified G protein was essentially devoid of internal viral proteins. Microdilution plates were coated with purified G protein, and sera from humans vaccinated against rabies were tested for the presence of antibodies. Results were compared with those of the rapid fluorescent focus inhibition assay, which is the standard neutralization assay for antibodies to rabies virus. The results of this comparison indicate that the enzyme-linked immunosorbent assay for G is a reliable and simple alternative to the neutralization test.
狂犬病病毒的包膜糖蛋白G可诱导中和抗体，这些抗体对于预防狂犬病很重要。该蛋白从纯化病毒中溶解出来，通过差速离心和蔗糖密度梯度离心，再经高效液相色谱法进行分离。利用免疫印迹和酶联免疫吸附测定技术对G蛋白的溶解和纯化条件进行了优化。与传统抗血清和单克隆抗体的反应表明，纯化的G蛋白基本不含病毒内部蛋白。将纯化的G蛋白包被在微量稀释板上，检测接种过狂犬病疫苗的人的血清中抗体的存在情况。将结果与快速荧光灶抑制试验的结果进行比较，快速荧光灶抑制试验是检测狂犬病病毒抗体的标准中和试验。比较结果表明，针对G蛋白的酶联免疫吸附测定是一种可靠且简便的中和试验替代方法。
