Disposable platform for bacterial lysis and nucleic acid amplification based on a single USB-powered printed circuit board.
Disposable platform for bacterial lysis and nucleic acid amplification based on a single USB-powered printed circuit board.
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DOI：
10.1371/journal.pone.0284424
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发表时间：
2023
期刊：
影响因子：
3.7
通讯作者：
Yager, Paul
中科院分区：
文献类型：
--
作者：
Shah, Kamal G.;Roller, Mike;Kumar, Sujatha;Bennett, Steven;Heiniger, Erin;Looney, Katriel;Buser, Joshua;Bishop, Joshua D.;Yager, Paul
电子和微流体技术的最新进展使几个研究小组能够开发出完全集成的、样品到结果的等温核酸扩增测试（NAAT）平台。然而，高昂的组件数量和成本限制了这些平台在诊所以外的低资源环境（包括家庭）中的应用。许多NAAT包括基于柔性电路或多个印刷电路板（PCB）的复杂的多组件加热器电子器件，以支持基本的NAAT步骤，例如裂解、样品失活和核酸扩增。相比之下，目前用于家庭使用的商业测定，例如用于怀孕或排卵的那些包括电子设备，通常只有一个板载PCB。这项工作描述了一种可推广的策略，将所有加热器和控制它们所需的电子设备集成到单个低成本USB供电的PCB上。我们构建了一个应用这些原理的多路复用一次性NAAT（“MD NAAT”）平台，在同一PCB上集成了将小区域加热到接近沸腾的小面积加热器（用于病原体裂解和灭活）和大面积加热器（用于扩增）。我们表明，这两类加热器具有很高的板内和设备间的再现性，尽管只加热NAAT墨盒从下面。我们通过裂解耐甲氧西林金黄色葡萄球菌（MRSA）细胞验证了小面积加热器，并通过进行两种类型的等温NAAT（等温链置换扩增（iSDA）和环介导等温扩增（LAMP））验证了大面积加热器。这些结果证明了将NAAT加热器和控制电子设备集成到单个印刷电路板上的优点，并且是将NAAT转化为家庭的一步。
Recent advances in electronics and microfluidics have enabled several research groups to develop fully integrated, sample-to-result isothermal nucleic acid amplification test (NAAT) platforms for the point of care. However, high component counts and costs have limited translation of these platforms beyond the clinic to low-resource settings—including homes. Many NAATs include complex, multi-component heater electronics based on flex circuits or multiple printed circuit boards (PCBs) to support essential NAAT steps such as lysis, sample deactivation, and nucleic acid amplification. In contrast, current commercial assays for home use, such as those for pregnancy or ovulation that include electronics, typically have just one onboard PCB. This work describes a generalizable strategy to integrate all heaters and the electronics needed to control them onto a single low-cost, USB-powered PCB. We built a multiplexable disposable NAAT (“MD NAAT”) platform that applies these principles, integrating small-area heaters that heat small regions to near-boiling (for pathogen lysis and deactivation) and large-area heaters (for amplification) on the same PCB. We show that both classes of heaters have high intra-board and inter-device reproducibility despite only heating a NAAT cartridge from below. We validated the small-area heaters by lysing methicillin-resistant Staphylococcus aureus (MRSA) cells and the large-area heaters by performing two types of isothermal NAATs (isothermal strand displacement amplification (iSDA) and loop-mediated isothermal amplification (LAMP)). These results demonstrate the merit of integrating NAAT heaters and control electronics onto a single printed circuit board and are a step toward translating NAATs to the home.
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影响因子：
3.7
作者：
Pardy, Tamas;Tulp, Indrek;Stewart, Ray
通讯作者：
Stewart, Ray
DOI：
10.1016/s0140-6736(17)32152-9
发表时间：
2017-09-16
期刊：
Lancet (London, England)
影响因子：
--
作者：
GBD 2016 Causes of Death Collaborators
通讯作者：
GBD 2016 Causes of Death Collaborators
影响因子：
6.1
作者：
Oblath EA;Henley WH;Alarie JP;Ramsey JM
通讯作者：
Ramsey JM
影响因子：
7.4
作者：
Huang S;Abe K;Bennett S;Liang T;Ladd PD;Yokobe L;Anderson CE;Shah K;Bishop J;Purfield M;Kauffman PC;Paul S;Welch AE;Strelitz B;Follmer K;Pullar K;Sanchez-Erebia L;Gerth-Guyette E;Domingo G;Klein E;Englund JA;Fu E;Yager P
通讯作者：
Yager P
影响因子：
4.2
作者：
Toley, Bhushan J.;Covelli, Isabela;Yagera, Paul
通讯作者：
Yagera, Paul