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用于通过环介导等温扩增检测传染性微生物的坚固且低成本的开源设备。

Robust and low-cost open-source device for detecting infectious microorganisms by loop-mediated isothermal amplification.

作者信息

Otero Jorge, Rodríguez-Lázaro Miguel A, Martínez-Trejo Arturo, Mbanze Daniel, Solana Gorka, Vergara Andrea, Bosch Salvador, Gozal David, Vila Jordi, Farré Ramon

机构信息

Unit of Biophysics and Bioengineering, School of Medicine and Health Sciences, University of Barcelona. Casanova 143, 08036 Barcelona, Spain.

CIBER de Enfermedades Respiratorias. Monforte de Lemos 3-5, 28029 Madrid, Spain.

出版信息

HardwareX. 2024 Aug 24;19:e00568. doi: 10.1016/j.ohx.2024.e00568. eCollection 2024 Sep.

Abstract

Loop-Mediated Isothermal Amplification (LAMP) is a useful technique for detecting infectious microorganisms in human fluids since it performs similarly to conventional PCR, the results are obtained faster and no thermocyclers or complex devices are required. Since only two isothermal blocks (95 °C to lyse cells and 65 °C for DNA amplification) are needed, LAMP is particularly suited for applications in Low- and Middle-Income Countries (LMICs). To validate such assumption, we first designed and tested Arduino-controlled LAMP thermoblocks to process a considerable number of samples simultaneously with a low-energy consumption to enable routine use under worst-case conditions (no main power source and low ambient temperatures). The thermoblocks were tested when battery-powered at temperature down to 5 °C, showing high stability in well temperatures (<0.8 °C). The charge required for both thermoblocks to simultaneously achieve the target temperatures after switching on and to keep their working temperatures were 4.1 A·h and 2.4 A·h/h, respectively. Second, we implemented a low-cost viewer with LEDs and filters to detect the fluorescent LAMP reaction. All the components required for the instrument are for general purpose and readily available by e-commerce. Thus, the LAMP device allows for considerable autonomy by using a typical car battery in rural and itinerant healthcare or field hospitals in LMICs, even under difficult environmental conditions.

摘要

环介导等温扩增技术(LAMP)是一种用于检测人体体液中感染性微生物的实用技术,因为它的性能与传统聚合酶链反应(PCR)相似,能更快获得结果,且无需热循环仪或复杂设备。由于仅需两个等温模块(95°C用于细胞裂解,65°C用于DNA扩增),LAMP特别适用于低收入和中等收入国家(LMICs)的应用。为验证这一假设,我们首先设计并测试了由 Arduino 控制的 LAMP 热模块,以便在低能耗情况下同时处理大量样本,从而能够在最坏情况条件下(无主电源和低环境温度)实现常规使用。当电池供电且温度低至5°C时对热模块进行测试,结果显示孔内温度具有高稳定性(<0.8°C)。两个热模块在开启后同时达到目标温度并保持其工作温度所需的电量分别为4.1 A·h和2.4 A·h/h。其次,我们采用了一种带有发光二极管(LED)和滤光片的低成本观察器来检测荧光LAMP反应。该仪器所需的所有组件均为通用组件,可通过电子商务轻松获取。因此,即使在困难的环境条件下,LAMP设备通过在农村和巡回医疗保健场所或LMICs的野战医院使用典型的汽车电池,也能实现相当程度的自主运行。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0dfd/11405915/adef4c85dc69/ga1.jpg

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