摘要
将大肠杆菌表达的GII-4型诺如病毒(Norovirus,NV)衣壳蛋白免疫新西兰大白兔,制备多克隆抗体;收集抗血清,酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测其效价,并对经聚合酶链式反应法(polymerase chain reaction,PCR)验证为阳性的NV感染样品进行检测,比较两种检测方法的差异性。以原核表达的诺如病毒衣壳蛋白作为抗原制备的NV特异性多克隆抗体具有良好的免疫反应性,抗体效价为1∶10000,可以用于免疫学检测。
New Zealand rabbits were immunized with the recombinant norovirus(NV) capsid protein expressed in E. coli to prepare polyclonal antibody. The antiserum was collected and evaluated by ELISA method, and a comparative evaluation with PCR was carried out by using both methods to detect NV-positive samples identified by PCR. The antiserum had a good immunoreactivity at a dilution of 1:10 000, suggesting its availability for immunological experiments. Compared with PCR, the ELISA method was more simple and rapid. Although the sensitivity of ELISA was worse than that of PCR, this problem was able to be meliorated by purified virus. This study establishes the basis for a rapid and economical norovirus enzyme-linked immunosorbent assay kit.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2014年第11期105-108,共4页
Food Science
基金
国家自然科学基金面上项目(31071525)
山东省“泰山学者”建设工程专项
教育部“长江学者和创新团队发展计划”项目(IRT1188)
国家自然科学基金青年科学基金项目(31101281)
关键词
诺如病毒
衣壳蛋白
原核表达
抗血清
norovirus
capsid protein
prokaryotic expression system
polyclonal antibody
