Equipment-free nucleic acid extraction and amplification on a simple paper disc for point-of-care diagnosis of rotavirus A
Equipment-free nucleic acid extraction and amplification on a simple paper disc for point-of-care diagnosis of rotavirus A
复制标题
在简单纸盘上进行无设备核酸提取和扩增，用于轮状病毒 A 的现场诊断
DOI：
10.1016/j.aca.2018.02.068
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发表时间：
2018-08-14
影响因子：
6.2
通讯作者：
Kong, Jilie
中科院分区：
文献类型：
--
作者：
Ye, Xin;Xu, Jin;Kong, Jilie
在临床诊断中使用基于纸张的方法是一个迅速发展的研究课题，吸引了人们的极大兴趣。一些小组试图在单个微流控纸片上实现集成的核酸测试，包括提取、放大和读出功能。然而，这些研究无法克服复杂的改装和制造要求、漫长的周转时间，或者对泵、热循环器或离心机等复杂设备的需求。在这里，我们报告了一种非常简单的纸质测试，用于轮状病毒A的护理诊断，轮状病毒A是导致儿童胃肠炎的最常见病原体之一。这种纸基检测方法可以在5分钟内完成核酸的提取，然后用25分钟的时间来扩增目标序列，结果是肉眼立即可以看到的，或者通过UV-Vis吸光度进行定量。这种低成本的方法不需要额外的设备，并且无论是在实验室还是在护理地点都很容易使用。轮状病毒A的检测限为1×10(3)拷贝/毫升。此外，对48份临床粪便标本进行检测，灵敏度和特异度均达到100%。综上所述，本纸质测试满足了医疗保健点诊断工具的主要要求，并具有应用于疾病预防、控制和精确诊断的潜力。(C)2018爱思唯尔B.V.保留所有权利。
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 x 10(3) 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. (c) 2018 Elsevier B.V. All rights reserved.