PATH, Seattle, Washington, United States of America.
PLoS One. 2011 May 9;6(5):e19738. doi: 10.1371/journal.pone.0019738.
Molecular assays targeted to nucleic acid (NA) markers are becoming increasingly important to medical diagnostics. However, these are typically confined to wealthy, developed countries; or, to the national reference laboratories of developing-world countries. There are many infectious diseases that are endemic in low-resource settings (LRS) where the lack of simple, instrument-free, NA diagnostic tests is a critical barrier to timely treatment. One of the primary barriers to the practicality and availability of NA assays in LRS has been the complexity and power requirements of polymerase chain reaction (PCR) instrumentation (another is sample preparation).
METHODOLOGY/PRINCIPAL FINDINGS: In this article, we investigate the hypothesis that an electricity-free heater based on exothermic chemical reactions and engineered phase change materials can successfully incubate isothermal NA amplification assays. We assess the heater's equivalence to commercially available PCR instruments through the characterization of the temperature profiles produced, and a minimal method comparison. Versions of the prototype for several different isothermal techniques are presented.
CONCLUSIONS/SIGNIFICANCE: We demonstrate that an electricity-free heater based on exothermic chemical reactions and engineered phase change materials can successfully incubate isothermal NA amplification assays, and that the results of those assays are not significantly different from ones incubated in parallel in commercially available PCR instruments. These results clearly suggest the potential of the non-instrumented nucleic acid amplification (NINA) heater for molecular diagnostics in LRS. When combined with other innovations in development that eliminate power requirements for sample preparation, cold reagent storage, and readout, the NINA heater will comprise part of a kit that should enable electricity-free NA testing for many important analytes.
针对核酸 (NA) 标志物的分子检测方法对于医学诊断变得越来越重要。然而，这些方法通常局限于富裕的发达国家，或者局限于发展中国家的国家参考实验室。在资源匮乏的环境中（LRS）存在许多地方性传染病，这些地区缺乏简单、无仪器、NA 诊断检测方法，这是及时治疗的关键障碍。在 LRS 中实现 NA 检测的实用性和可用性的主要障碍之一是聚合酶链反应（PCR）仪器的复杂性和电力要求（另一个是样品制备）。
方法/主要发现：在本文中，我们研究了这样一个假设，即基于放热化学反应和工程相变材料的无电加热器可以成功孵育等温 NA 扩增检测。我们通过对产生的温度曲线进行特征描述和最小方法比较，评估了该加热器与市售 PCR 仪器的等效性。介绍了几种不同等温技术的原型版本。
结论/意义：我们证明了基于放热化学反应和工程相变材料的无电加热器可以成功孵育等温 NA 扩增检测，并且那些在市售 PCR 仪器中平行孵育的检测结果没有显著差异。这些结果清楚地表明了无仪器核酸扩增（NINA）加热器在 LRS 中用于分子诊断的潜力。当与其他开发中的创新相结合，这些创新消除了对样品制备、冷试剂储存和读出的电力需求时，NINA 加热器将成为一个试剂盒的一部分，该试剂盒应能够实现许多重要分析物的无电 NA 检测。
