Enzyme-linked immunosorbent assays using virus-like particles containing mutations of conserved residues on envelope protein can distinguish three flavivirus infections.
Enzyme-linked immunosorbent assays using virus-like particles containing mutations of conserved residues on envelope protein can distinguish three flavivirus infections.
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使用含有包膜蛋白保守残基突变的病毒样颗粒进行酶联免疫吸附测定，可以区分三种黄病毒感染。
DOI：
10.1080/22221751.2020.1797540
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发表时间：
2020
影响因子：
13.2
通讯作者：
Wang,Wei-Kung
中科院分区：
文献类型：
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作者：
Tsai,Wen-Yang;Driesse,Kaitlin;Tsai,Jih-Jin;Hsieh,Szu-Chia;SznajderGranat,Robert;Jenkins,Olivia;Chang,Gwong-Jen;Wang,Wei-Kung
寨卡病毒(ZIKV)最近在黄病毒流行地区的暴发突显了进行敏感和特定的血清学测试的必要性。以前我们和其他人报道了登革病毒(DENV)囊膜蛋白上的关键融合环(FL)残基和/或BC环(BCL)残基，这些残基可以被黄病毒交叉反应的人单抗和多克隆血清识别。为了提高ZIKV的血清学诊断水平，我们用ZIKV、DENV1和西尼罗病毒(WNV)的野生型(WT)和FL或FL/BCL变异病毒样颗粒(VLP)进行了酶联免疫吸附试验(EL ISA)，并对不同感染ZIKV、DENV和WNV的逆转录聚合酶链式反应(RT-PCR)确诊病例的恢复期血清或血浆标本进行了检测。ZIKV WT-VLP的敏感性为100%，特异性为52.9%，FL/bc1突变体VLP的敏感性和特异性分别为83.3%和92.2%。同样，DENV1和WNV WT-VLP的灵敏度/特异度分别为100%/70.0%和100%/56.3%，FL突变体VLP的特异度分别提高到93.3%和83.0%。对于IgM-EL ISA，ZIKV、DENV1和WNV WT-VLP对原发感染的特异度分别为96.4%、92.3%和91.4%，FL或FL/bclVLP的特异度提高到93.7-99.3%。提出了一种基于突变型和WT-VLP免疫球蛋白结合的算法，用于区分ZIKV、DENV和WNV的原发感染以及DENV和ZIKV继发感染和既往的DENV感染，这可能是更好地了解多种黄病毒共存地区ZIKV的血清阳性率和致病机制的有力工具。
The recent outbreaks of Zika virus (ZIKV) in flavivirus-endemic regions highlight the need for sensitive and specific serological tests. Previously we and others reported key fusion loop (FL) residues and/or BC loop (BCL) residues on dengue virus (DENV) envelope protein recognized by flavivirus cross-reactive human monoclonal antibodies and polyclonal sera. To improve ZIKV serodiagnosis, we employed wild type (WT) and FL or FL/BCL mutant virus-like particles (VLP) of ZIKV, DENV1 and West Nile virus (WNV) in enzyme linked immunosorbent assays (ELISA), and tested convalescent-phase serum or plasma samples from reverse-transcription PCR-confirmed cases with different ZIKV, DENV and WNV infections. For IgG ELISA, ZIKV WT-VLP had a sensitivity of 100% and specificity of 52.9%, which was improved to 83.3% by FL/BCL mutant VLP and 92.2% by the ratio of relative optical density of mutant to WT VLP. Similarly, DENV1 and WNV WT-VLP had a sensitivity/specificity of 100%/70.0% and 100%/56.3%, respectively; the specificity was improved to 93.3% and 83.0% by FL mutant VLP. For IgM ELISA, ZIKV, DENV1 and WNV WT-VLP had a specificity of 96.4%, 92.3% and 91.4%, respectively, for primary infection; the specificity was improved to 93.7–99.3% by FL or FL/BCL mutant VLP. An algorithm based on a combination of mutant and WT-VLP IgG ELISA is proposed to discriminate primary ZIKV, DENV and WNV infections as well as secondary DENV and ZIKV infection with previous DENV infections; this could be a powerful tool to better understand the seroprevalence and pathogenesis of ZIKV in regions where multiple flaviviruses co-circulate.