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A simple, robust and equipment-free DNA amplification readout in less than 30 seconds

机译：在不到30秒的情况下，在不到30秒内的简单，稳健和无设备的DNA放大读数

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摘要

Molecular based diagnostic methods rely on the amplification of pathogen DNA but naked eye visualization of results is still challenging. We present here a simple and highly reliable DNA amplification readout system for naked eye detection of isothermally or PCR amplified DNA in less than 30 seconds. This system utilizes spermine to precipitate DNA amplicons and initiate bridging flocculation of a mix of charcoal and diatomaceous earth particles in suspension. In the absence of amplification, the charcoal particles remain suspended resulting in a black, non-transparent colloid solution while positive samples in which DNA amplification has occurred can be identified within seconds as the particles flocculate and settle leaving a transparent liquid phase. We have coupled this method with our rapid dipstick DNA purification method and isothermal DNA amplification to create a simple four-step diagnostic system that can be preassembled to reduce unnecessary manipulation in the field. The method's simplicity, low cost, minimal equipment and clear presence/absence readout makes it ideal for rapid diagnostic testing in the laboratory and in situations where users have limited technical training or resources including high school science classes and field-based research.
机译：基于分子基诊断方法依赖于病原体DNA的扩增，但肉眼可视化的结果仍然挑战。我们在这里介绍一种简单且高度可靠的DNA放大读数读数系统，用于肉眼检测等温或PCR扩增DNA，在不到30秒的时间内。该系统利用精霉素沉淀DNA扩增子，并引发悬浮液中木炭和硅藻土颗粒混合物的桥接絮凝。在没有扩增的情况下，木炭颗粒保持悬浮，得到黑色，非透明胶体溶液，而可以在颗粒絮凝并沉降留下透明液相时在几秒钟内鉴定其中发生DNA扩增的正样品。我们已经通过快速的Dipstick DNA净化方法和等温DNA放大器耦合了这种方法，以创造一个简单的四步诊断系统，可以预告以减少该领域的不必要的操纵。该方法的简单性，低成本，最小的设备和清晰的存在/缺失读数使其成为实验室中快速诊断测试的理想选择，并且在用户有限的技术培训或资源包括高中科学课程和基于实地的研究中的情况下。

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