Enzyme-Linked Immunosorbent Assay for Detection of Filovirus Species-Specific Antibodies
Enzyme-Linked Immunosorbent Assay for Detection of Filovirus Species-Specific Antibodies
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DOI：
10.1128/cvi.00170-10
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发表时间：
2010-11-01
影响因子：
--
通讯作者：
Takada, Ayato
中科院分区：
文献类型：
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作者：
Nakayama, Eri;Yokoyama, Ayaka;Takada, Ayato
已经开发了几种用于检测丝状病毒特异性抗体的酶联免疫吸附测定（ELISA）。然而，区分丝状病毒各种特异性抗体的诊断方法（为血清学分类提供基础）尚不容易获得。我们建立了一种ELISA，使用His标记的分泌形式的跨膜糖蛋白（GP）的五个不同的埃博拉病毒（EBOV）物种和一个马尔堡病毒（MARV）株作为抗原的丝状病毒物种特异性抗体的检测。通过测试从用病毒样颗粒免疫的小鼠以及从感染EBOV或MARV的人和非人灵长类动物收集的抗血清来评估基于GP的ELISA。在我们的ELISA中，在大多数小鼠抗血清中观察到IgG抗体的交叉反应性很小。尽管来自一些患者和猴的血清和血浆显示出与来自多种丝状病毒种属的GP的显著交叉反应性，但针对与感染个体的病毒种属同源的GP抗原一致检测到IgG的最高反应。我们进一步证实，MARV特异性IgM抗体在感染急性期从患者采集的标本中特异性检测到。这些结果证明了我们的ELISA用于诊断以及生态和血清调查研究的有用性。
Several enzyme-linked immunosorbent assays (ELISAs) for the detection of filovirus-specific antibodies have been developed. However, diagnostic methods to distinguish antibodies specific to the respective species of filoviruses, which provide the basis for serological classification, are not readily available. We established an ELISA using His-tagged secreted forms of the transmembrane glycoproteins (GPs) of five different Ebola virus (EBOV) species and one Marburg virus (MARV) strain as antigens for the detection of filovirus species-specific antibodies. The GP-based ELISA was evaluated by testing antisera collected from mice immunized with virus-like particles as well as from humans and nonhuman primates infected with EBOV or MARV. In our ELISA, little cross-reactivity of IgG antibodies was observed in most of the mouse antisera. Although sera and plasma from some patients and monkeys showed notable cross-reactivity with the GPs from multiple filovirus species, the highest reactions of IgG were uniformly detected against the GP antigen homologous to the virus species that infected individuals. We further confirmed that MARV-specific IgM antibodies were specifically detected in specimens collected from patients during the acute phase of infection. These results demonstrate the usefulness of our ELISA for diagnostics as well as ecological and serosurvey studies.