The recent Zika virus (ZIKV) outbreak demonstrates that cost-effective clinical diagnostics are urgently needed to detect and distinguish viral infections to improve patient care. Unlike dengue virus (DENV), ZIKV infections during pregnancy correlate with severe birth defects, including microcephaly and neurological disorders. Because ZIKV and DENV are related flaviviruses, their homologous proteins and nucleic acids can cause cross-reactions and false-positive results in molecular, antigenic, and serologic diagnostics. We report the characterization of monoclonal antibody pairs that have been translated into rapid immunochromatography tests to specifically detect the viral nonstructural 1 (NS1) protein antigen and distinguish the four DENV serotypes (DENV1–4) and ZIKV without cross-reaction. To complement visual test analysis and remove user subjectivity in reading test results, we used image processing and data analysis for data capture and test result quantification. Using a 30-μl serum sample, the sensitivity and specificity values of the DENV1–4 tests and the pan-DENV test, which detects all four dengue serotypes, ranged from 0.76 to 1.00. Sensitivity/specificity for the ZIKV rapid test was 0.81/0.86, respectively, using a 150-μl serum input. Serum ZIKV NS1 protein concentrations were about 10-fold lower than corresponding DENV NS1 concentrations in infected patients; moreover, ZIKV NS1 protein was not detected in polymerase chain reaction–positive patient urine samples. Our rapid immunochromatography approach and reagents have immediate application in differential clinical diagnosis of acute ZIKV and DENV cases, and the platform can be applied toward developing rapid antigen diagnostics for emerging viruses.

最近的寨卡病毒 (ZIKV) 爆发表明，迫切需要具有成本效益的临床诊断来检测和区分病毒感染，以改善患者护理。与登革热病毒 (DENV) 不同，怀孕期间的 ZIKV 感染与严重的出生缺陷相关，包括小头畸形和神经系统疾病。由于 ZIKV 和 DENV 是相关的黄病毒，它们的同源蛋白和核酸在分子、抗原和血清学诊断中会引起交叉反应和假阳性结果。我们报告了已转化为快速免疫色谱测试的单克隆抗体对的特征，以特异性检测病毒非结构 1 (NS1) 蛋白抗原并区分四种 DENV 血清型 (DENV1-4) 和 ZIKV，而不会发生交叉反应。为了补充视觉测试分析并消除用户在阅读测试结果时的主观性，我们使用图像处理和数据分析进行数据捕获和测试结果量化。使用 30 μl 血清样本，DENV1-4 检测和检测所有四种登革热血清型的 pan-DENV 检测的敏感性和特异性值范围为 0.76 至 1.00。ZIKV 快速测试的敏感性/特异性分别为 0.81/0.86，使用 150-μl 血清输入。血清 ZIKV NS1 蛋白浓度比感染患者相应的 DENV NS1 浓度低约 10 倍；此外，在聚合酶链反应阳性的患者尿液样本中未检测到 ZIKV NS1 蛋白。我们的快速免疫层析方法和试剂可立即应用于急性 ZIKV 和 DENV 病例的临床鉴别诊断，