三种抗原检测H_1N_1猪流感病毒抗体的比较  

Comparation of Three Methods for Detection of Antibodies to H_1N_1 Swine Influenza Virus

作　　者：杨倩[1] 薛春宜[1] 朱道中[2] 李晓明[1] 曹永长[1] 

机构地区：[1]中山大学生命科学学院有害生物控制与资源利用国家重点实验室,广东广州510006 [2]广东出入境检验检疫局检验检疫技术中心,广东广州510623

出　　处：《广东畜牧兽医科技》2014年第1期16-20,共5页Guangdong Journal of Animal and Veterinary Science

基　　金：国家质检总局科技计划项目(2011IK004)

摘　　要：本研究采用昆虫杆状病毒表达系统制备了H1N1亚型猪流感病毒的HA蛋白、类病毒脂质体、病毒样颗粒,分别作为抗原进行包被,采用间接酶联免疫吸附实验(ELISA)检测H1N1猪流感病毒抗体。特异性实验结果表明,单独表达的HA蛋白和类病毒脂质体特异性良好,但是病毒样颗粒不能区分不同亚型的流感病毒;敏感性实验结果表明,类病毒脂质体作为抗原的敏感性最好;重复性实验结果表明,三种抗原的重复性好,ELISA实验的批间和批内变异系数均小于10%;稳定性实验结果表明,在4℃可至少稳定保存1年。综上所述,类病毒脂质体有较高的免疫反应性,与灭活全病毒相比有更好的安全性,在抗体水平测定方面有较好的应用前景。HA protein, virus-like liposomes, and virus-like particles, expressed by Bac-to-Bac baculovirus expression system, were coated as antigens to detect the H1N1 swine influenza virus antibody using indirect enzyme-linked immunosorbent assay （ELISA）. The results of specificity experiments showed that the single-expressed HA protein and virus-like liposomes had good specificity, but the virus-like particles could not distinguish different subtypes of influenza virus. The results of sensitivity experiments showed that virus-like liposomes had the best sensitivity. The results of repetitive experiments showed that the repetitiveness of the three methods was high, and the coefficient of variation within-rim and between-run were less than 1%. The results of stability experiments showed that three antigens could be stable at 4 ~C for at least one year. In a word, the virus-like liposome has strongest immunoreactivity, and is safer than the inactivated whole influenza virus. Thus it represents a promising method in detection of H1N1 antibodies.

关 键 词：昆虫杆状病毒表达系统 猪流感病毒 类病毒脂质体 抗体检测 

分 类 号：S852.43[农业科学—基础兽医学]