Alleviating the "Sample to Sequence" Bottleneck Using Novel Microfluidic Lab-on-a-Chip Nucleic Acid Extraction Technologies

这项研究将基于英国和日本现有的技术和专业知识，开发和评估一个新的系统，这将改善我们基于物种特异性基因序列检测、测量和研究海洋生物的方式。目前，鉴定、枚举和分析海洋基因序列的最佳方法依赖于收集水样，这些水样被送回资源丰富的集中实验室，由训练有素的技术人员对其进行处理和分析。这需要时间，延迟了潜在的重要结果（例如有害物种的存在和数量），而且成本高昂，限制了可以处理的样本数量，最终降低了我们监测海洋生物的分辨率。随着海洋对不断变化的气候和人为影响作出反应，这一点现在比以往任何时候都更加重要。在这项工作中，一个关键的限制步骤是从样本中去除遗传物质的过程，无论是整个细胞、生物体还是它们的残留物，并将其净化到可以精确测量的程度；“提取瓶颈”。现有的自动化样品处理机器人通常体积庞大，复杂，耗电，价格昂贵，而且没有广泛使用。微流控芯片实验室（LOC）技术减少了传统上在实验室工作台上进行的分析过程的规模。例如，微型管道（通常直径为十分之一毫米）与微型泵、阀门和光学装置一起用于采集样品，并与一套试剂一起操作样品，以便在一小部分时间内以最小的样品/化学品消耗和机器人方式进行相对复杂的实验室过程，从而避免了对专家的需求。在这个项目中，我们将利用LOC技术的优势，通过一种新型设备来解决提取瓶颈，该设备将与“前端”采样器和“后端”分析仪接口，形成一个集成的遗传传感器平台。这将适用于对人类健康、海洋生态和以海洋为中心的工业具有高度重要性的一系列目标生物的检测和定量。该项目将证明将LOC基因提取与现有采样器和分析相结合可以显着提高分辨率和易用性，从而我们可以监测基本的生物变量。

This research will develop and evaluate a new system, based on pre-existing technology and expertise within the UK and Japan, which will improve the way in which we can detect, measure and study ocean biology based on species-specific genetic sequences. Current, best methods for the identification, enumeration and analysis of genetic sequences in the ocean rely upon the collection of water samples, which are returned to a centralized and highly resourced laboratory where they are processed and analyzed by highly trained technical staff. This takes time, delaying potentially important results (e.g. the presence and quantity of harmful species), and is expensive, limiting the number of samples that can be processed and ultimately reducing the resolution with which we can monitor ocean biology. This is now more important than ever as the oceans respond to changing climatic and anthropogenic influences. A key limiting step in this endeavor is the process of removing genetic material from the sample, whether it be whole cells, organisms or their remnants, and purifying it to the point at which it can be measured accurately; the 'extraction bottleneck.' Existing, automated sample processing robots are typically bulky, complicated, power hungry, prohibitively expensive, and not widely available. Microfluidic Lab on a Chip (LOC) technologies reduce the scale of analytical processes traditionally performed on a lab bench. For example, miniature pipes (typically one tenth of a millimetre across) together with miniature pumps, valves and optics are used to take in sample, and manipulate it along with a suite of reagents to undertake a relatively complex laboratory process in a fraction of the time with minimal sample / chemical consumption and robotically, thus obviating the need for a specialist. In this project, we will capitalize upon the advantages of LOC technology to address the extraction bottleneck with a novel device that will interface with 'front-end' samplers and 'back-end' analyzers to form an integrated, genetic sensor platform. This will be tailored for the detection and quantification of a range of target organisms of high importance to human health, ocean ecology and ocean-centric industries. The project will demonstrate proof of concept that the integration of LOC genetic extraction with existing samplers and analytics can significantly improve the resolution and ease with which we can monitor fundamental biological variables.