Competitive detection of influenza neutralizing antibodies using a novel bivalent fluorescence-based microneutralization assay (BiFMA)
Competitive detection of influenza neutralizing antibodies using a novel bivalent fluorescence-based microneutralization assay (BiFMA)
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DOI：
10.1016/j.vaccine.2015.05.049
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发表时间：
2015-07-09
期刊：
影响因子：
5.5
通讯作者：
Martinez-Sobrido, Luis
中科院分区：
文献类型：
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作者：
Baker, Steven F.;Nogales, Aitor;Martinez-Sobrido, Luis
禽源性甲型流感人畜共患病受到密切监测，可能是具有大流行潜力的病毒株的迹象。成功的疫苗接种和人类甲型流感病毒的恢复通常导致可以中和病毒感染的抗体的诱导。为了改进用于检测中和抗体的长期和新一代方法，我们采用了一种新的基于抗体的方法，该方法允许在单个样品中进行多种抗原检测。该方法的核心是单循环感染性甲型流感病毒（sciIAV），其中功能性血凝素（HA）基因被改变为编码绿色或单体红色荧光蛋白（分别为GFP和mRFP），HA由稳定的HA表达细胞系反式补充。通过使用具有非重叠发射光谱的荧光蛋白，这种新型的基于二价荧光的微量中和测定（BiFMA）可用于在单一反应中检测针对两种不同流感分离株的中和抗体，使实验速度加倍，同时使所需血清量减半。此外，该方法可用于快速鉴定流感广泛中和抗体。重要的是，这种新型BiFMA可用于BSL-2设施下的任何给定流感HA假型病毒，包括高致病性流感HA分离株。(C)2015爱思唯尔有限公司版权所有。
Avian-derived influenza A zoonoses are closely monitored and may be an indication of virus strains with pandemic potential. Both successful vaccination and convalescence of influenza A virus in humans typically results in the induction of antibodies that can neutralize viral infection. To improve long-standing and new-generation methodologies for detection of neutralizing antibodies, we have employed a novel reporter-based approach that allows for multiple antigenic testing within a single sample. Central to this approach is a single-cycle infectious influenza A virus (sciIAV), where a functional hemagglutinin (HA) gene was changed to encode either the green or the monomeric red fluorescent protein (GFP and mRFP, respectively) and HA is complemented in trans by stable HA-expressing cell lines. By using fluorescent proteins with non-overlapping emission spectra, this novel bivalent fluorescence-based microneutralization assay (BiFMA) can be used to detect neutralizing antibodies against two distinct influenza isolates in a single reaction, doubling the speed of experimentation while halving the amount of sera required. Moreover, this approach can be used for the rapid identification of influenza broadly neutralizing antibodies. Importantly, this novel BiFMA can be used for any given influenza HA-pseudotyped virus under BSL-2 facilities, including highly pathogenic influenza HA isolates. (C) 2015 Elsevier Ltd. All rights reserved.