摘要
初步研究六种致病性白蛉病毒原核表达核蛋白(Nucleocapsid protein,NP)抗原的抗原性,为开发相应的血清学诊断试剂提供理论依据。利用原核表达系统表达纯化六种致病性白蛉病毒核蛋白,并分别免疫新西兰白兔。通过间接ELISA方法及Western-blotting方法对所获得的兔免疫血清进行交叉反应检测,确定其抗体ELISA滴度。此外,分别用上述六种致病性白蛉病毒NP抗原包被ELISA反应板并对发热伴血小板减少综合征病毒(Severe fever with thrombocytopenia syndrome virus,SFTSV)病人血清进行间接ELISA检测,确定其血清学交叉反应及IgG抗体滴度。通过原核表达系统表达并纯化的六种白蛉病毒NP抗原浓度及纯度均达到免疫和检测要求。ELISA及Western-blotting检测结果显示六种病毒NP抗原兔免疫血清中的每种血清与其余五种病毒NP抗原可能存在血清学交叉反应。SFTSV病人血清与除SFTSV-NP以外的五种NP抗原亦可能存在部分交叉反应。本研究初步评价了原核表达的六种致病性白蛉病毒核蛋白的抗原性和免疫反应性,为相应诊断试剂的研制奠定了基础。
This article aimed to study the antigenicity of nucleocapsid proteins（NPs）in six pathogenic phleboviruses and to provide theoretical evidence for the development of serological diagnostic reagents.NPs of six pathogenic phleboviruses were expressed and purified using aprokaryotic expression system and rabbits were immunized with individual recombinant NPs.Cross-reactions among NPs and rabbit sera were determined by both indirect ELISA and Western blotting analyses,and the sera titer was determined by indirect ELISA.Furthermore,sera from SFTS patients were also detected by each recombinant NP as a coating antigen using indirect ELISA.The cross-reactions and the sera titer were subsequently determined.Both the concentration and purity of recombinant NPs of six pathogenic phleboviruses met the standards for immunization and detection.The results of indirect ELISA and Western blotting showed that each anti-phlebovirus NP rabbit immune serum had potential serological cross-reactivity with the other five virus NP antigens.Furthermore,the sera from SFTS patients also had cross-reactivity with the other five NP antigens to a certain extent.Our preliminary study evaluated the antigenicity and immune reactivity of six pathogenic phleboviruses NPs and laid the foundation for the development of diagnostic reagents.
出处
《病毒学报》
CAS
CSCD
北大核心
2014年第4期387-390,共4页
Chinese Journal of Virology
基金
艾滋病和病毒性肝炎等重大传染病防治项目:重大传染病应急处置检测技术平台(2013ZX10004-101)
