利用类病毒脂质体抗原检测H5N1亚型禽流感病毒抗体ELISA方法的建立  

Establishment of Indirect Enzyme-linked Immunosorbent Assay for Detecting H5N1 Avian Influenza Virus Antibody Based on Virus-like Liposome

作　　者：杨倩[1,2] 朱道中[3] 薛春宜[1] 李晓明[1] 曹永长[1] 段燕喻[3] 王莹[3] 吴晓薇[3] 刘志玲[3] 

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]中国疾病预防控制中心病毒病预防控制所,北京102206 [3]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：《动物医学进展》2015年第1期5-10,共6页Progress In Veterinary Medicine

基　　金：国家质检总局科技计划项目(2011IK004)

摘　　要：利用昆虫杆状病毒表达系统制备了H5N1亚型禽流感病毒(AIV)HA蛋白、类病毒脂质体和病毒样颗粒,分别作为包被抗原,建立了相应的检测H5N1亚型禽流感病毒抗体的ELISA方法。特异性试验、敏感性试验、重复性试验和稳定性试验结果表明,利用3种抗原包被所建立的ELISA方法均具有良好的重复性和稳定性,批间和批内变异系数均小于10%,但单独表达的HA蛋白和类病毒脂质体特异性更好,而且类病毒脂质体有更高的免疫反应性,与灭活全病毒相比安全性更高,故在H5N1亚型AIV抗体水平检测方面更具有应用前景。HA protein,virus-like liposomes and virus-like particles,expressed by Bac-to-Bac baculovirus expression system,were coated respectively as antigens to establish an indirect enzyme-linked immunosor-bent assay （ELISA） for detecting AIV H5N1 antibody.The results of specificity test,sensitivity test,re-petitivity test and stability test showed that the three established ELISAs had good repeatability and stabil-ity,and the coefficient of variation within-run and between-run were less than 10%.The results of specific-ity test indicated the single-expressed HA protein and virus-like liposomes had good specificity,but the ELISA based on the virus-like particles can not distinguish different subtypes of influenza virus.In addi-tion,the virus-like liposome has strong immunoreactivity,and it is safer than the inactivated avian influen-za virus.Therefore,the virus-like liposome represents to be more promising than the others in detection of antibodies.

关 键 词：昆虫杆状病毒表达系统 禽流感病毒 类病毒脂质体 抗体检测 

分 类 号：S852.659.5[农业科学—基础兽医学]