A new competitive ELISA detects West Nile virus infection using monoclonal antibodies against the precursor-membrane protein of West Nile virus
A new competitive ELISA detects West Nile virus infection using monoclonal antibodies against the precursor-membrane protein of West Nile virus
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DOI：
10.1016/j.jviromet.2012.12.002
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发表时间：
2013-03-01
影响因子：
3.1
通讯作者：
Shimizu, Shinya
中科院分区：
文献类型：
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作者：
Hirota, Jiro;Shimizu, Shinya
前体膜蛋白（prM）存在于未成熟的西尼罗病毒（WNV）颗粒的包膜中。抗prM抗体存在于黄病毒感染的动物血清中，并且它们被称为相对病毒物种特异性抗体。然而，目前还没有已知的WNV特异性表位阻断或竞争性酶联免疫吸附试验（c-ELISA）检测抗prM抗体的报道。制备了两种抗WNV-prM单克隆抗体（SHW-29 C2和SHW-3182），并使用这些单克隆抗体开发了c-ELISA。使用WNV感染的鸡血清评价c-ELISA。这两种c-ELISA检测抗prM抗体在WNV感染的鸡血清中，并表现出对日本脑炎病毒，圣路易斯脑炎病毒，和墨累谷脑炎病毒的抗血清几乎没有交叉反应。SHW-29 C2型和SHW-3182型c-ELISA对WNV感染后3周鸡血清的平均抑制率分别为61.6%和71.8%。在c-ELISA中的抑制百分比与IgG间接ELISA的光密度值之间观察到高度相关性。此外，基于SHW-3182的c-ELISA检测针对多种WNV毒株的抗体。用c-ELISA检测抗PrM抗体可用于西尼罗河病毒的血清学诊断。皇冠版权所有（C）2012由Elsevier B. V.出版。保留所有权利。
Precursor membrane protein (prM) is found in the envelope of immature West Nile virus (WNV) particles. Anti-prM antibodies are found in flavivirus-infected animal sera, and they are known as relatively virus species specific antibodies. However, there are no known reports of WNV-specific epitope blocking or competitive enzyme-linked immunosorbent assay (c-ELISA) that detect anti-prM antibodies. Two anti-WNV-prM monoclonal antibodies (SHW-29C2 and SHW-3182) were generated, and c-ELISAs were developed using these monoclonal antibodies. The c-ELISAs were evaluated using WNV-infected chicken sera. Both c-ELISAs detected anti-prM antibodies in WNV-infected chicken sera and showed little cross-reactivity to antisera against Japanese encephalitis virus, St. Louis encephalitis virus, and Murray valley encephalitis virus. The average inhibition of chicken sera at 3 weeks post WNV infection was 61.6% in SHW-29C2-based c-ELISA and 71.8% in SHW-3182-based c-ELISA. High correlation was seen between percent inhibition in the c-ELISAs and optical density values of an IgG indirect ELISA. Additionally, SHW-3182-based c-ELISA detected antibodies against a wide variety of WNV strains. Detecting anti-PrM antibodies using c-ELISA could be useful for WNV serodiagnosis. Crown Copyright (C) 2012 Published by Elsevier B.V. All rights reserved.