The use of paper-based methods for clinical diagnostics is a rapidly expanding research topic attracting a great deal of interest. Some groups have attempted to realize an integrated nucleic acid test on a single microfluidic paper chip, including extraction, amplification, and readout functions. However, these studies were not able to overcome complex modification and fabrication requirements, long turn-around times, or the need for sophisticated equipment like pumps, thermal cyclers, or centrifuges. Here, we report an extremely simple paper-based test for the point-of-care diagnosis of rotavirus A, one of the most common pathogens that causes pediatric gastroenteritis. This paper-based test could perform nucleic acid extraction within 5 min, then took 25 min to amplify the target sequence, and the result was visible to the naked eye immediately afterward or quantitative by the UV-Vis absorbance. This low-cost method does not require extra equipment and is easy to use either in a lab or at the point-of-care. The detection limit for rotavirus A was found to be 1 × 103 copies/mL. In addition, 100% sensitivity and specificity were achieved when testing 48 clinical stool samples. In conclusion, the present paper-based test fulfills the main requirements for a point-of-care diagnostic tool, and has the potential to be applied to disease prevention, control, and precision diagnosis.

中文翻译：

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在简单的纸盘上进行无设备核酸提取和扩增，用于轮状病毒 A 的即时诊断

使用基于纸张的方法进行临床诊断是一个迅速扩展的研究课题，引起了极大的兴趣。一些团队试图在单个微流控纸芯片上实现整合核酸检测，包括提取、扩增和读出功能。然而，这些研究无法克服复杂的修改和制造要求、较长的周转时间或对泵、热循环仪或离心机等复杂设备的需求。在这里，我们报告了一种极其简单的纸质测试，用于轮状病毒 A 的即时诊断，轮状病毒 A 是引起小儿胃肠炎的最常见病原体之一。这种纸质测试可以在5分钟内完成核酸提取，然后用25分钟来扩增目标序列，结果立即肉眼可见或通过UV-Vis吸光度定量。这种低成本的方法不需要额外的设备，并且易于在实验室或床旁使用。轮状病毒 A 的检测限为 1 × 103 拷贝/mL。此外，在测试 48 个临床粪便样本时实现了 100% 的灵敏度和特异性。总之，目前的纸质测试满足了即时诊断工具的主要要求，具有应用于疾病预防、控制和精确诊断的潜力。测试 48 个临床粪便样本时实现了 100% 的灵敏度和特异性。总之，目前的纸质测试满足了即时诊断工具的主要要求，具有应用于疾病预防、控制和精确诊断的潜力。测试 48 个临床粪便样本时实现了 100% 的灵敏度和特异性。总之，目前的纸质测试满足了即时诊断工具的主要要求，具有应用于疾病预防、控制和精确诊断的潜力。