Several tropical fever viruses transmitted by mosquitoes including zika, dengue, and chikungunya, are becoming a serious problem in global public health. Simple diagnostic tools in early stages are strongly required to monitor and prevent these diseases. Paper diagnostic platforms can provide a solution for these needs, with integration of fluidic control techniques and isothermal amplification methods. Here, we demonstrate a Lab-on-paper for all-in-one molecular diagnostics of zika, dengue, and chikungunya virus from human serum. The entire process of nucleic acid testing that involves sampling, extraction, amplification, and detection is simply operated on a single paper chip. Based on the engineered structure of paper materials and dried chemicals on the all-in-one chip, serum samples containing the target virus RNA were simply added by automatic flow from distilled water injection. Target RNA molecules were concentrated on the binding pad with chitosan and then transported to reaction pads following a pH increase for specific reverse transcription loop-mediated isothermal amplification with fluorescence signal generation. Three targets, zika virus, dengue virus, and chikungunya virus, in human serum were simultaneously detected on the all-in-one paper chip within 60 min at 65 °C. The all-in-one paper chip can be used as a real-time quantitative assay for 5–5000 copies of zika virus RNA. This all-in-one device was successfully used with 5 clinical specimens of zika and dengue virus from real patients. We believe that the proposed all-in-one paper chip can provide a portable, low-cost, user-friendly, sensitive, and specific NAT platform with great potential in point-of-care diagnostics.

蚊子传播的几种热带热病毒包括寨卡病毒，登革热和基孔肯雅热，正在成为全球公共卫生中的一个严重问题。早期需要使用简单的诊断工具来监视和预防这些疾病。纸质诊断平台可以通过集成流体控制技术和等温扩增方法来提供满足这些需求的解决方案。在这里，我们演示了一种纸上实验室，可对寨卡病毒，登革热和基孔肯雅热进行多合一分子诊断来自人血清的病毒。涉及采样，提取，扩增和检测的整个核酸测试过程都可以在单个纸屑上简单地操作。基于纸质材料和干燥化学药品在多合一芯片上的工程结构，只需从蒸馏水注入中自动流加含有目标病毒RNA的血清样品即可。将目标RNA分子浓缩在带有壳聚糖的结合垫上，然后在pH升高后转运至反应垫，以进行特定的逆转录环介导的等温扩增，并产生荧光信号。寨卡病毒，登革热病毒和基孔肯雅病毒这三个目标，在65°C下60分钟内在多合一纸屑芯片上同时检测到人血清中的TNF-α。多合一纸屑芯片可用于实时定量测定5至5000个寨卡病毒RNA。这种多合一的设备已成功用于实际患者的5个寨卡病毒和登革热临床标本。我们相信，提出的多合一纸质芯片可以提供便携式，低成本，用户友好，灵敏且特定的NAT平台，在即时诊断方面具有巨大潜力。