Diagnosis of Zika Virus Infection by Peptide Array and Enzyme-Linked Immunosorbent Assay.
Diagnosis of Zika Virus Infection by Peptide Array and Enzyme-Linked Immunosorbent Assay.
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DOI：
10.1128/mbio.00095-18
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发表时间：
2018-03-06
期刊：
影响因子：
6.4
通讯作者：
Lipkin WI
中科院分区：
文献类型：
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作者：
Mishra N;Caciula A;Price A;Thakkar R;Ng J;Chauhan LV;Jain K;Che X;Espinosa DA;Montoya Cruz M;Balmaseda A;Sullivan EH;Patel JJ;Jarman RG;Rakeman JL;Egan CT;Reusken CBEM;Koopmans MPG;Harris E;Tokarz R;Briese T;Lipkin WI
寨卡病毒（ZIKV）与胎儿死产、小头畸形、颅内钙化和受感染母亲垂直传播后的眼部异常有关。在成人中，感染可能引发自身免疫性炎症性多发性神经病。传播最常见的是被感染的伊蚊叮咬，但也可能通过性交或接受血液制品发生。通过检测病毒RNA，可以在发病后10天内在血清或血浆中，在发病后3周内在全血中，以及在精液中长达3个月内进行连续诊断。尽管如此，血清学诊断仍然至关重要，因为很少有患者在感染的急性期获得分子诊断，并且感染可能仅与不提示分子检测的轻度或不明显的疾病相关。血清学诊断被免疫血清与ZIKV最近出现的地区流行的其他黄病毒的交叉反应性所混淆。因此，我们建立了一个高密度的微阵列，包括非冗余的12聚体肽，瓷砖，与一个残基重叠，寨卡病毒，登革热，黄热病，西尼罗河，伊尔赫斯，奥罗波什，和基孔肯雅病毒的蛋白质组。血清学分析使得能够发现具有高灵敏度的ZIKV NS 2B 20-残基肽（96.0%）和特异性（95.9%）与自然感染或接种登革热、基孔肯雅热、黄热病、西尼罗河、蜱传脑炎，或日本脑炎病毒在微阵列测定和早期恢复期血清的酶联免疫吸附测定（ELISA（出现症状感染后2至3周）。寨卡病毒（ZIKV）作为一种致畸剂的出现对全球公共卫生构成了深刻挑战。当患者出现病毒血症时，感染的分子诊断在3周内是直接的。然而，此后对历史暴露的血清学诊断受到交叉反应性的混淆。使用高密度肽阵列，平铺黄病毒的选择的蛋白质组，以确定ZIKV特异性肽，我们建立了两个测定，使敏感和特异性诊断暴露于ZIKV。这些测定可用于指导处于潜在暴露于ZIKV的风险中的母亲的临床管理，并能够深入了解ZIKV感染的流行病学。
Zika virus (ZIKV) is implicated in fetal stillbirth, microcephaly, intracranial calcifications, and ocular anomalies following vertical transmission from infected mothers. In adults, infection may trigger autoimmune inflammatory polyneuropathy. Transmission most commonly follows the bite of infected Aedes mosquitoes but may also occur through sexual intercourse or receipt of blood products. Definitive diagnosis through detection of viral RNA is possible in serum or plasma within 10 days of disease onset, in whole blood within 3 weeks of onset, and in semen for up to 3 months. Serological diagnosis is nonetheless critical because few patients have access to molecular diagnostics during the acute phase of infection and infection may be associated with only mild or inapparent disease that does not prompt molecular testing. Serological diagnosis is confounded by cross-reactivity of immune sera with other flaviviruses endemic in the areas where ZIKV has recently emerged. Accordingly, we built a high-density microarray comprising nonredundant 12-mer peptides that tile, with one-residue overlap, the proteomes of Zika, dengue, yellow fever, West Nile, Ilheus, Oropouche, and chikungunya viruses. Serological analysis enabled discovery of a ZIKV NS2B 20-residue peptide that had high sensitivity (96.0%) and specificity (95.9%) versus natural infection with or vaccination against dengue, chikungunya, yellow fever, West Nile, tick-borne encephalitis, or Japanese encephalitis virus in a microarray assay and an enzyme-linked immunosorbent assay (ELISA) of early-convalescent-phase sera (2 to 3 weeks after onset of symptomatic infection). The emergence of Zika virus (ZIKV) as a teratogen is a profound challenge to global public health. Molecular diagnosis of infection is straightforward during the 3-week period when patients are viremic. However, serological diagnosis thereafter of historical exposure has been confounded by cross-reactivity. Using high-density peptide arrays that tile the proteomes of a selection of flaviviruses to identify a ZIKV-specific peptide, we established two assays that enable sensitive and specific diagnosis of exposure to ZIKV. These assays may be useful in guiding clinical management of mothers at risk for potential exposure to ZIKV and enable insights into the epidemiology of ZIKV infections.