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Development of a Flow-Free Automated Colorimetric Detection Assay Integrated with Smartphone for Zika NS1

机译:与智能手机集成的Zika NS1无流量自动比色检测分析方法的开发

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摘要

The Zika virus (ZIKV) is an emerging flavivirus transmitted to humans by mosquitoes that can potentially cause microcephaly, Guillain–Barré Syndrome, and other birth defects. Effective vaccines for Zika have not yet been developed. There is a necessity to establish an easily deployable, high-throughput, low-cost, and disposable point-of-care (POC) diagnostic platform for ZIKV infections. We report here an automated magnetic actuation platform suitable for a POC microfluidic sandwich enzyme-linked immunosorbent assay (ELISA) using antibody-coated superparamagnetic beads. The smartphone integrated immunoassay is developed for colorimetric detection of ZIKV nonstructural protein 1 (NS1) antigen using disposable chips to accommodate the reactions inside the chip in microliter volumes. An in-house-built magnetic actuator platform automatically moves the magnetic beads through different aqueous phases. The assay requires a total of 9 min to automatically control the post-capture washing, horseradish peroxidase (HRP) conjugated secondary antibody probing, washing again, and, finally, color development. By measuring the saturation intensity of the developed color from the smartphone captured video, the presented assay provides high sensitivity with a detection limit of 62.5 ng/mL in whole plasma. These results advocate a great promise that the platform would be useful for the POC diagnosis of Zika virus infection in patients and can be used in resource-limited settings.
机译:寨卡病毒(ZIKV)是一种新兴的黄病毒,是通过蚊子传播给人类的,可能引起小头畸形,格林-巴利综合症和其他先天性缺陷。尚未开发出用于寨卡的有效疫苗。必须为ZIKV感染建立易于部署,高吞吐量,低成本和一次性的即时护理(POC)诊断平台。我们在这里报告了一个自动的磁激励平台,适用于使用抗体包被的超顺磁珠的POC微流体夹心酶联免疫吸附测定(ELISA)。开发了智能手机集成免疫测定法,用于使用一次性芯片对ZIKV非结构蛋白1(NS1)抗原进行比色检测,以适应微升体积内芯片内的反应。内置的磁性执行器平台可自动将磁珠移动通过不同的水相。该测定总共需要9分钟,以自动控制捕获后的洗涤,辣根过氧化物酶(HRP)偶联的二抗探测,再次洗涤以及最终显色。通过从智能手机捕获的视频中测量显色的饱和度强度,本测定法可提供高灵敏度,整个血浆中的检测限为62.5 ng / mL。这些结果表明,该平台可用于患者Zika病毒感染的POC诊断,并可以在资源有限的环境中使用。

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