Next Generation of Ultra-Sensitive DNA Diagnostics for Gastric Infections

下一代胃感染超灵敏 DNA 诊断

• 批准号: 1811634
• 金额: --
• 依托单位: University of Strathclyde
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1811634
• 关键词: Next; Generation; Ultra; Sensitive; DNA

We have been working on methodologies to detect DNA sequences relating to infectious disease for a number of years with a focus on using functionalised nanoparticles and surface enhanced Raman scattering. This work has produced a number of different assay approaches which are now under development through Renishaw Diagnostics Ltd. Now that these assays are moving into the hands of the end user, the market feedback suggests that improvements can be made to the assays to make them more competitive with the latest developments in the sector and also to improve the ease of use for non-specialists. This PhD studentship is designed to investigate the detection of specific DNA sequences indicative of gastric infections in a multiplexed and simple manner without the need for PCR amplification of the targets as is used in a majority of commercially available assays. The challenge of moving to a PCR free based assay is mainly in terms of the sensitivity requirements where PCR greatly increases the chance of detection but also in terms of the simplicity of hybridisation of the target with the probes through the control of the length of the DNA using PCR The aim of this proposal is to develop a new assay to detect DNA sequences in a clinically relevant range (less than 107 copies) for gastric infections without any enzyme steps. The objectives are:1. To investigate the use of a stationary phase such as nitrocellulose for the immobilisation of a probe sequence of DNA to capture the target DNA and wash away excess. This will also involve the addition of DNA functionalised nanoparticles which we will use to hybridise to the captured target sequence on the surface. We will take a further leap in sensitivity by using a nanoparticle assembly approach which we have already shown to work through DNA hybridisation. This will take place only on the nanoparticles attached to the DNA sequence on the surface.2. Having tested the above system we will move towards clinical samples and evaluate the performance of this assay format with clinical samples which contain a multitude of components not found in the model test systems.3. To create a working device for verification and validation of this approach for use in ultrasensitive DNA detection which should be possible in less than 1 hour from sample collection.

多年来，我们一直致力于研究检测与传染病相关的DNA序列的方法，重点是使用功能化纳米颗粒和表面增强拉曼散射。这项工作已经产生了许多不同的检测方法，目前正在通过雷尼绍诊断有限公司进行开发。现在，这些检测方法正在进入最终用户的手中，市场反馈表明，可以对检测方法进行改进，使其在该行业的最新发展中更具竞争力，并提高非专业人员的易用性。该博士研究生项目旨在研究以多重和简单的方式检测指示胃感染的特定DNA序列，而不需要像大多数市售检测中使用的那样对靶点进行PCR扩增。转向无PCR检测的挑战主要在于灵敏度要求，其中PCR大大增加了检测的机会，而且还在于通过使用PCR控制DNA的长度使靶与探针杂交的简单性。本提案的目的是开发一种新的检测方法，以检测临床相关范围内的DNA序列。（少于107个拷贝）用于胃感染，而没有任何酶步骤。目标是：1.研究使用固定相（如硝酸纤维素）固定DNA探针序列，以捕获靶DNA并洗去过量DNA。这还将涉及添加DNA功能化纳米颗粒，我们将使用这些纳米颗粒与表面上捕获的靶序列杂交。我们将通过使用纳米颗粒组装方法在灵敏度上取得进一步的飞跃，我们已经证明了这种方法可以通过DNA杂交来发挥作用。这将只发生在纳米颗粒上附着到表面上的DNA序列。2.在测试了上述系统之后，我们将转向临床样本，并使用包含大量在模型测试系统中未发现的组分的临床样本来评估该测定格式的性能。创建一个工作设备，用于验证和确认这种方法，用于超灵敏的DNA检测，这应该是可能的，在不到1小时的样品收集。