A Rapid and Low-Cost PCR Thermal Cycler for Infectious Disease Diagnostics
A Rapid and Low-Cost PCR Thermal Cycler for Infectious Disease Diagnostics
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DOI：
10.1371/journal.pone.0149150
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发表时间：
2016-02-12
期刊：
影响因子：
3.7
通讯作者：
Wong, Season S.
中科院分区：
文献类型：
--
作者：
Chan, Kamfai;Wong, Pui-Yan;Wong, Season S.
能够以便携、低成本的形式广泛提供传染病的快速诊断，将标志着全球卫生领域向前迈出了一大步。许多分子诊断方法是基于利用热循环仪分别进行聚合酶链式反应(PCR)和逆转录聚合酶链式反应(RT-PCR)来扩增和检测DNA和RNA。遗憾的是，大多数商用热循环器价格昂贵，需要持续供电，因此不适合在资源较少的环境中使用。我们之前已经报道了一种使用真空绝缘不锈钢热瓶食品罐头扩增DNA的低成本和简单的方法，我们将其命名为热瓶热循环器或TTC。在这里，我们描述了一种改进的设置的使用，以实现通过逆转录聚合酶链式反应(RT-PCR)检测病毒RNA靶标，从而扩展了TTC在低资源环境下识别高传染性、基于RNA病毒的疾病的能力。TTC成功地展示了对性传播疾病、艾滋病毒/艾滋病、埃博拉出血热和登革热的DNA或RNA目标的快速和灵敏的检测。我们的创新TTC的建造成本不到200美元，并且至少有8根管子的容量。在速度方面，TTC的性能超过了测试的商用热循环器。当TTC与低成本的终点检测技术相结合时，如核酸侧向流动分析或基于手机的荧光检测器，TTC将在低资源环境下增加现场分子诊断的可用性。
The ability to make rapid diagnosis of infectious diseases broadly available in a portable, low-cost format would mark a great step forward in global health. Many molecular diagnostic assays are developed based on using thermal cyclers to carry out polymerase chain reaction (PCR) and reverse-transcription PCR for DNA and RNA amplification and detection, respectively. Unfortunately, most commercial thermal cyclers are expensive and need continuous electrical power supply, so they are not suitable for uses in low-resource settings. We have previously reported a low-cost and simple approach to amplify DNA using vacuum insulated stainless steel thermoses food cans, which we have named it thermos thermal cycler or TTC. Here, we describe the use of an improved set up to enable the detection of viral RNA targets by reverse-transcription PCR (RT-PCR), thus expanding the TTC's ability to identify highly infectious, RNA virus-based diseases in low resource settings. The TTC was successful in demonstrating high-speed and sensitive detection of DNA or RNA targets of sexually transmitted diseases, HIV/AIDS, Ebola hemorrhagic fever, and dengue fever. Our innovative TTC costs less than $200 to build and has a capacity of at least eight tubes. In terms of speed, the TTC's performance exceeded that of commercial thermal cyclers tested. When coupled with low-cost endpoint detection technologies such as nucleic acid lateral-flow assay or a cell-phone-based fluorescence detector, the TTC will increase the availability of on-site molecular diagnostics in low-resource settings.