Use of recombinant E protein domain III-based enzyme-linked immunosorbent assays for differentiation of tick-borne encephalitis serocomplex flaviviruses from mosquito-borne flaviviruses
Use of recombinant E protein domain III-based enzyme-linked immunosorbent assays for differentiation of tick-borne encephalitis serocomplex flaviviruses from mosquito-borne flaviviruses
复制标题
DOI：
10.1128/jcm.42.9.4101-4110.2004
复制
发表时间：
2004-09-01
影响因子：
9.4
通讯作者：
Barrett, ADT
中科院分区：
文献类型：
--
作者：
Holbrook, MR;Shope, RE;Barrett, ADT
黄病毒感染的血清学诊断由于黄病毒交叉反应抗体的存在而变得复杂，所述黄病毒交叉反应抗体产生黄病毒感染的假阳性结果，特别是在多于一种病毒流行的地区。目前蜱传黄病毒感染的诊断试剂已被发现与黄热病或登革热病毒阳性血清交叉反应。本研究利用重组黄病毒E蛋白结构域3（rE-D3）作为诊断试剂，以区分蚊子和蜱传播的黄病毒感染。这项研究发现，使用rE-D3酶联免疫吸附测定（ELISA）为基础的格式允许蚊子或蜱传黄病毒特异性血清之间的区别，但不是蜱传脑炎（TBE）黄病毒血清复合体的成员。在Western印迹和ELISA中，发现抗几种TBE血清复合物rE-D3衍生的血清与TBE血清复合物内的异源rE-D3交叉反应，但不与来自蚊媒黄病毒的血清交叉反应。还发现针对TBE血清复合物病毒产生的小鼠超免疫血清与TBE血清复合物rE-D3特异性反应，但不与来自蚊媒病毒的rE-D3反应，反之亦然。当使用病毒衍生抗原进行类似试验时，观察到特异性和灵敏度均丧失。这些结果表明黄病毒rE-D3将是用于检测TBE血清复合物黄病毒感染的有用试剂，其中几种是潜在的生物威胁剂，但不能提供区分血清复合物的不同成员感染的能力。
The serological diagnosis of infection by flaviviruses is complicated by the presence of flavivirus crossreactive antibodies that produce false-positive results for flavivirus infections, especially in regions where more than one virus is endemic. Current diagnostic reagents for tick-borne flavivirus infection have been found to cross-react with yellow fever- or dengue virus-positive sera. This study utilized recombinant flavivirus E protein domain 3 (rE-D3) as a diagnostic reagent to differentiate between infection by mosquito- and tick-borne flaviviruses. This study found that the use of rE-D3 in an enzyme-linked immunosorbent assay (ELISA)-based format allowed the differentiation between serum specific for either mosquito- or tick-borne flaviviruses, but not among the members of the tick-borne encephalitis (TBE) serocomplex of flaviviruses. Sera derived against several TBE serocomplex rE-D3 were found to cross-react with heterologous rE-D3 within the TBE serocomplex, but not with those from mosquito-borne flaviviruses, in both Western blots and ELISAs. Mouse hyperimmune sera generated against TBE serocomplex viruses were also found to react specifically with TBE serocomplex rE-D3, but not with rE-D3 from mosquito-borne viruses and vice versa. When a similar test using virus-derived antigen was performed, a loss of both specificity and sensitivity was observed. These results indicate that flavivirus rE-D3 would be a useful reagent for the detection of infection by TBE serocomplex flaviviruses, several of which are potential biothreat agents, but would not provide the ability to differentiate among infections by separate members of the serocomplex.