Moving toward rapid and low-cost point-of-care molecular diagnostics with a repurposed 3D printer and RPA.
Moving toward rapid and low-cost point-of-care molecular diagnostics with a repurposed 3D printer and RPA.
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DOI：
10.1016/j.ab.2018.01.008
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发表时间：
2018-03-15
影响因子：
2.9
通讯作者：
Wong S
中科院分区：
文献类型：
--
作者：
Chan K;Wong PY;Parikh C;Wong S
传统上，大多数基于核酸扩增的分子诊断测试都是在集中设置中完成的。近年来，已开发出在远离中心实验室的资源匮乏环境中使用的即时护理检测。虽然大多数专家一致认为，即时护理分子检测是非常需要的，但它们作为具有成本效益和易于操作的检测方法的可用性仍然是一个未实现的目标。在本文中，我们讨论了我们的努力，以开发重组酶聚合酶扩增反应为基础的测试，将满足这些标准。首先，我们描述了我们在重新利用低成本3D打印机作为一个平台，可以进行中通量，快速，高性能的核酸提取的努力。接下来，我们将讨论如何使用3D打印机的加热床或我们开发的解构的低成本热循环器快速放大和分析这些纯化的模板。在这两种方法中，模板DNA或RNA的实时等温扩增和检测都可以使用低成本的便携式检测器或智能手机摄像头完成。最后，我们展示了我们使用食源性病原体和寨卡病毒的技术的能力。我们的低成本方法不使用复杂和高成本的组件，使其适合资源有限的环境。整合和商业化后，它将提供简单的从样本到答案的分子诊断。
Traditionally, the majority of nucleic acid amplification-based molecular diagnostic tests are done in centralized settings. In recent years, point-of-care tests have been developed for use in low-resource settings away from central laboratories. While most experts agree that point-of-care molecular tests are greatly needed, their availability as cost-effective and easy-to-operate tests remains an unmet goal. In this article, we discuss our efforts to develop a recombinase polymerase amplification reaction-based test that will meet these criteria. First, we describe our efforts in repurposing a low-cost 3D printer as a platform that can carry out medium-throughput, rapid, and high-performing nucleic acid extraction. Next, we address how these purified templates can be rapidly amplified and analyzed using the 3D printer’s heated bed or the deconstructed, low-cost thermal cycler we have developed. In both approaches, real-time isothermal amplification and detection of template DNA or RNA can be accomplished using a low-cost portable detector or smartphone camera. Last, we demonstrate the capability of our technologies using foodborne pathogens and the Zika virus. Our low-cost approach does not employ complicated and high-cost components, making it suitable for resource-limited settings. When integrated and commercialized, it will offer simple sample-to-answer molecular diagnostics.
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影响因子：
3.9
作者：
Linnes JC;Fan A;Rodriguez NM;Lemieux B;Kong H;Klapperich CM
通讯作者：
Klapperich CM
使用磁珠和液滴阵列平行提取 RNA。
影响因子：
6.1
作者：
Shi X;Chen CH;Gao W;Chao SH;Meldrum DR
通讯作者：
Meldrum DR
影响因子：
7.4
作者：
Connelly, John T.;Rolland, Jason P.;Whitesides, George M.
通讯作者：
Whitesides, George M.
影响因子：
3.7
作者：
Chan, Kamfai;Wong, Pui-Yan;Wong, Season S.
通讯作者：
Wong, Season S.
影响因子：
3.7
作者：
Lillis L;Lehman D;Singhal MC;Cantera J;Singleton J;Labarre P;Toyama A;Piepenburg O;Parker M;Wood R;Overbaugh J;Boyle DS
通讯作者：
Boyle DS