Measuring Antibody Neutralization of Dengue Virus (DENV) Using a Flow Cytometry-Based Technique
Measuring Antibody Neutralization of Dengue Virus (DENV) Using a Flow Cytometry-Based Technique
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DOI：
10.1007/978-1-4939-0348-1_3
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发表时间：
2014-01-01
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影响因子：
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通讯作者：
de Silva, Aravinda M.
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文献类型：
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作者：
de Alwis, Ruklanthi;de Silva, Aravinda M.
登革病毒（DENV）是一种新出现的病毒，威胁着世界三分之二以上的人口。通过血清学对登革热感染的特异性诊断是基于检测DENV特异性抗体（包括中和抗体（Abs））的测定。中和抗体是一个重要的，如果不是主要的，保护机制，从自然登革病毒（DENV）感染以及。目前用于测量针对DENV的中和Ab应答的金标准测定是空斑减少中和测定（PRNT）。然而，该测定缓慢且费力，并且利用生理学上不相关的细胞系。在这里，我们描述了一种相对高通量的，基于流式细胞术的DENV中和试验，该试验已被优化用于人单核细胞悬浮细胞系U937 + DC-SIGN或更常用的贴壁猴肾细胞Vero-81。
Dengue virus (DENV) is an emerging virus that threatens over two-third of the world's population. The specific diagnosis of dengue infection by serology is based on assays that detect DENV-specific antibodies including neutralizing antibodies (Abs). Neutralizing Abs are an important, if not the main, mechanism of protection from natural dengue virus (DENV) infection as well. The current gold-standard assay for measuring neutralizing Ab responses against DENV is the plaque reduction neutralization assay (PRNT). However, this assay is slow and laborious and utilizes physiologically irrelevant cell lines. Here, we describe a relatively high-throughput, flow cytometry-based neutralization assay for DENV that has been optimized for use with a human monocytic suspension cell line, U937 + DC-SIGN, or the more commonly used adherent monkey kidney cells, Vero-81.