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一种用于快速检测环境中大肠杆菌的可现场部署的低成本聚合酶链式反应(FLC-PCR)热循环仪。

A Field-Deployable and Low-Cost PCR (FLC-PCR) Thermocycler for the Rapid Detection of Environmental E. coli.

作者信息

Ferguson James, Duran Jesse, Killinen Wesley, Wagner Jason, Kulesza Caroline, Chatterley Christie, Li Yiyan

出版信息

Annu Int Conf IEEE Eng Med Biol Soc. 2020 Jul;2020:2209-2212. doi: 10.1109/EMBC44109.2020.9176519.

Abstract

This is a proof-of-concept study for the development of a field-deployable and low-cost PCR thermocycler (FLC-PCR) to perform Polymerase Chain Reaction (PCR) for the rapid detection of environmental E. coli. Four efficient (77.1 W) peltier modules are used as the central temperature control unit. One 250 W silicone heating pad is used for the heating lid. The PID (proportional-integral-derivative) control algorithm for the thermocycles is implemented by a low-cost 8-bit, 16 MHz microcontroller (ATMEGA328P-PU). ybbW and uidA genes from specific E. coli colonies were used as amplicons for the PCR reactions that were carried out by a commercial PCR machine (Bio-Rad) and our FLC-PCR thermocycler. The heating and cooling speeds averaged 1.11 ± 0.33°C/s which is on a par with the commercial bench-top PCR thermocycler and the efficiency of the heating lid outperformed the Bio-Rad PCR thermocycler. The overall cost of the system is lower than $200 which is more than ten times lower than commercially available units. The heating block can be customized to accommodate different PCR tubes and even microfluidic chambers. An 8000 W portable power generator will be used as the power supply for field studies.

摘要

这是一项概念验证研究,旨在开发一种可现场部署且低成本的聚合酶链反应(PCR)热循环仪(FLC-PCR),用于快速检测环境中的大肠杆菌。四个高效(77.1瓦)的珀尔帖模块用作中央温度控制单元。一个250瓦的硅胶加热垫用于加热盖。热循环的比例积分微分(PID)控制算法由一个低成本的8位、16兆赫兹微控制器(ATMEGA328P-PU)实现。来自特定大肠杆菌菌落的ybbW和uidA基因用作PCR反应的扩增子,这些反应由商用PCR仪(伯乐公司)和我们的FLC-PCR热循环仪进行。加热和冷却速度平均为1.11±0.33°C/秒,与商用台式PCR热循环仪相当,并且加热盖的效率优于伯乐公司的PCR热循环仪。该系统的总成本低于200美元,比市售设备低十多倍。加热块可定制,以容纳不同的PCR管甚至微流控腔室。一台功率为8000瓦的便携式发电机将用作现场研究的电源。

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