Next Generation of Ultra-Sensitive DNA Diagnostics for Gastric Infections

多年来，我们一直致力于研究检测与传染病有关的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，这些方法目前正在雷尼绍诊断有限公司开发中。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，使其与该领域的最新发展更具竞争力，并提高非专业人员使用的便利性。这个博士研究生的目的是研究以一种多路和简单的方式检测指示胃感染的特定DNA序列，而不需要像大多数商业分析中使用的那样对目标进行PCR扩增。搬到一个基于PCR免费化验的挑战主要是灵敏度的要求,PCR检测的机会大大增加而且简单的目标与探针的杂交通过DNA的长度的控制使用PCR这个建议的目的是开发一种新的试验来检测DNA序列在临床相关的范围内(少于107册)胃感染没有任何酶步骤。目标是：1。研究使用固定相（如硝化纤维素）固定探针DNA序列以捕获目标DNA并洗去多余的DNA。这也将涉及到DNA功能化纳米颗粒的添加，我们将使用它与表面捕获的目标序列杂交。我们将通过使用纳米粒子组装方法进一步提高灵敏度，我们已经证明了这种方法可以通过DNA杂交发挥作用。这只会发生在附着在表面DNA序列上的纳米颗粒上。在测试了上述系统之后，我们将转向临床样品，并使用包含模型测试系统中未发现的大量成分的临床样品评估该分析格式的性能。创建一个工作装置，用于验证和验证该方法，用于超灵敏DNA检测，该方法应该可以在样品收集后不到1小时内完成。

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.