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使用电池供电的薄膜电阻热循环仪进行快速DNA扩增。

Rapid DNA amplification using a battery-powered thin-film resistive thermocycler.

作者信息

Herold Keith E, Sergeev Nikolay, Matviyenko Andriy, Rasooly Avraham

机构信息

Fischell Department of Bioengineering, University of Maryland, College Park, MD, USA.

出版信息

Methods Mol Biol. 2009;504:441-58. doi: 10.1007/978-1-60327-569-9_24.

Abstract

A prototype handheld, compact, rapid thermocycler was developed for multiplex analysis of nucleic acids in an inexpensive, portable configuration. Instead of the commonly used Peltier heating/cooling element, electric thin-film resistive heater and a miniature fan enable rapid heating and cooling of glass capillaries leading to a simple, low-cost Thin-Film Resistive Thermocycler (TFRT). Computer-based pulse width modulation control yields heating rates of 6-7 K/s and cooling rates of 5 K/s. The four capillaries are closely coupled to the heater, resulting in low power consumption. The energy required by a nominal PCR cycle (20 s at each temperature) was found to be 57+/-2 J yielding an average power of approximately 1.0 W (not including the computer and the control system). Thus the device can be powered by a standard 9 V alkaline battery (or other 9 V power supply). The prototype TFRT was demonstrated (in a benchtop configuration) for detection of three important food pathogens (E. coli ETEC, Shigella dysenteriae, and Salmonella enterica). PCR amplicons were analyzed by gel electrophoresis. The 35 cycle PCR protocol using a single channel was completed in less then 18 min. Simple and efficient heating/cooling, low cost, rapid amplification, and low power consumption make the device suitable for portable DNA amplification applications including clinical point of care diagnostics and field use.

摘要

我们开发了一种原型手持式紧凑型快速热循环仪,用于以低成本、便携的配置对核酸进行多重分析。该热循环仪未采用常用的珀耳帖加热/冷却元件,而是使用电薄膜电阻加热器和微型风扇,实现了玻璃毛细管的快速加热和冷却,从而形成了一种简单、低成本的薄膜电阻热循环仪(TFRT)。基于计算机的脉宽调制控制实现了6 - 7 K/s的加热速率和5 K/s的冷却速率。四个毛细管与加热器紧密耦合,从而降低了功耗。一个标称PCR循环(每个温度下20秒)所需的能量为57±2焦耳,平均功率约为1.0瓦(不包括计算机和控制系统)。因此,该设备可以由标准的9伏碱性电池(或其他9伏电源)供电。已对原型TFRT(以台式配置)进行了演示,用于检测三种重要的食源性病原体(肠产毒性大肠杆菌、痢疾志贺氏菌和肠炎沙门氏菌)。通过凝胶电泳对PCR扩增产物进行分析。使用单通道的35个循环的PCR方案在不到18分钟内完成。简单高效的加热/冷却、低成本、快速扩增和低功耗使得该设备适用于便携式DNA扩增应用,包括临床即时诊断和现场使用。

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