Wadsworth Center, New York State Department of Health, Albany, New York, USA.
Wadsworth Center, New York State Department of Health, Albany, New York, USA.
EBioMedicine. 2017 Feb;16:136-140. doi: 10.1016/j.ebiom.2017.01.008. Epub 2017 Jan 10.
Rapid and accurate diagnosis of infectious agents is essential for patient care, disease control, and countermeasure development. The present serologic diagnosis of Zika virus (ZIKV) infection relies mainly on IgM-capture ELISA which is confounded with the flaw of cross-reactivity among different flaviviruses. In this communication, we report a multiplex microsphere immunoassay (MIA) that captures the diagnostic power of viral envelope protein (that elicits robust, yet cross-reactive antibodies to other flaviviruses) and the differential power of viral nonstructural proteins NS1 and NS5 (that induce more virus-type specific antibodies). Using 153 patient specimens with known ZIKV and/or dengue virus (DENV; a closely related flavivirus) infections, we showed that (i) ZIKV envelope-based MIA is equivalent or more sensitive than IgM-capture ELISA in diagnosing ZIKV infection, (ii) antibody responses to NS1 and NS5 proteins are more ZIKV-specific than antibody response to envelope protein, (iii) inclusion of NS1 and NS5 in the MIA improves the diagnostic accuracy when compared with the MIA that uses envelope protein alone. The multiplex MIA achieves a rapid diagnosis (turnaround time<4h) and requires small specimen volume (10μl) in a single reaction. This serologic assay could be developed for use in clinical diagnosis of ZIKV infection and for monitoring immune responses in vaccine trials.
快速准确地诊断病原体对于患者治疗、疾病控制和对策制定至关重要。目前，寨卡病毒（ZIKV）感染的血清学诊断主要依赖于 IgM 捕获 ELISA，该方法存在与其他黄病毒交叉反应的缺陷。在本研究中，我们报告了一种多重微球免疫分析（MIA），该方法结合了病毒包膜蛋白（诱导针对其他黄病毒的强烈但交叉反应性抗体）的诊断能力，以及病毒非结构蛋白 NS1 和 NS5 的差异能力（诱导更具病毒特异性的抗体）。使用 153 份已知感染 ZIKV 和/或登革热病毒（DENV；一种密切相关的黄病毒）的患者标本，我们表明：（i）基于 ZIKV 包膜的 MIA 在诊断 ZIKV 感染方面与 IgM 捕获 ELISA 等效或更敏感；（ii）与包膜蛋白相比，针对 NS1 和 NS5 蛋白的抗体反应更具 ZIKV 特异性；（iii）与仅使用包膜蛋白的 MIA 相比，将 NS1 和 NS5 纳入 MIA 可提高诊断准确性。该多重 MIA 可实现快速诊断（周转时间<4 小时），并且在单次反应中仅需要 10μl 的小样本量。这种血清学检测方法可用于临床诊断 ZIKV 感染，并用于监测疫苗试验中的免疫反应。
