Low cost nanowire diagnostic platform

实验室检测是准确诊断患者病情的重要组成部分，每年在英国进行约10亿次检测。随着对循证医学兴趣的扩大，实验室检查越来越多地用于根据需要定制个人治疗计划，监测疾病进展，分层风险和人群筛查计划。为了实现预测性、预防性和个性化医疗保健的常规应用，这些生化测试需要以低成本大规模进行，并且优选地还在需要的位置进行，而不是仅在临床实验室中进行。实现这一社会经济上高度理想的情况需要开发新的、更具成本效益的技术，用于量化分子，这些分子是诊断和管理特定疾病的生物标志物。该项目的目标是开发一种即时（point-of-care）纳米线诊断系统，以测量在一小滴血液中发现的炎症生物标志物，并建立在EPSRC纳米技术用于医疗保健项目EP/6061696/1的结果基础上。该项目将开发一种具有集成样品处理的低成本纳米线平台技术。纳米线制造将使用一种新颖的自上而下的工艺，温度低到足以在廉价的玻璃或聚合物衬底上进行大规模制造。我们的目标应用是呼吸系统疾病的诊断和管理，包括COPD（慢性阻塞性肺病）和哮喘，这些疾病通常会因病毒感染而加重。使用翻译方法，我们将专注于检测病毒感染和治疗的一些临床相关蛋白质生物标志物。病毒感染是一种免疫反应，其特征是抗病毒干扰素及其下游反应基因（如干扰素诱导蛋白10和β 2-微球蛋白）或激活标志物（如新蝶呤）的表达。这些可以用作感染的早期生物标志物，或用于评估或预测干扰素-α或β抗病毒治疗的临床反应，这些反应通常与中和抗体的检测结合测量。肿瘤坏死因子α（TNF α）和急性期蛋白C反应蛋白（CRP）也被用作与感染加重相关的炎症生物标志物。纳米线阵列将用于定量CRP、TNFa、新蝶呤、IP-10和β 2-微球蛋白的水平（用特异性抗体进行纳米线功能化）以及干扰素-β的抗体水平（用干扰素-β本身进行功能化）。该项目还将探索DNA适体作为合成抗体的潜力，这是一种比经典蛋白抗体更具成本效益和生物相容性的方法。许多疾病没有一个单一的生物标志物与疾病状态相关，因此需要使用一组生物标志物来更准确地诊断和管理疾病。该项目将拥有独特的临床样本-血清和诱导痰-从南安普顿总医院急性医疗单元收治的急性哮喘恶化患者中获得，其中60-80%是由呼吸道病毒引发的，并在其恢复后。我们还将获得哮喘志愿者的血清样本，这些志愿者正在接受吸入性干扰素-β的I期临床试验，该试验正在开发中，用于治疗病毒引起的哮喘急性发作。将结果与使用传统方法（ELISA）获得的数据和临床结局（例如呼吸道病毒检测、急性加重严重程度、治疗反应）进行直接比较。

Laboratory tests are a vital and growing part of the accurate diagnosis of a patient's condition, with around 1 billion tests performed in the UK each year. With the expansion of interest in evidence-based medicine, laboratory tests are increasingly used to tailor individual treatment plans according to need, to monitor disease progression, to stratify risk, and for population screening programs. To enable the routine application of predictive, preventative and personalized healthcare, these biochemical tests need to be performed at a large scale, at low cost, and preferably also at point-of-need locations rather than exclusively in clinical laboratories. The realization of this socio-economically highly desirable situationnecessitates the development of new, more cost-effective, technologies for the quantification of molecules that are biomarkers for the diagnosis and management of specific diseases.The project aim is to develop a point-of-care (PoC) nanowire diagnostic system to measure inflammatory biomarkers found in a small droplet of blood, and builds on results from EPSRC Nanotechnology for Healthcare project EP/6061696/1. The project will develop a low-cost nanowire platform technology with integrated sample processing. Nanowire fabrication will use a novel top down process and temperatures low enough for large scale manufacture on inexpensive glass or polymer substrates.Our target application is the diagnosis and management of respiratory diseases, including COPD (chronic obstructive pulmonary disease) and asthma, that are often exacerbated by viral infections. Using a translational approach, we will focus on the detection of a number of clinically relevant protein biomarkers of viral infection and treatment. Viral infectionstrigger an immune response characterized by expression of the antiviral interferons and their down stream response genes (e.g. interferon-inducible protein 10 and beta2-microglobulin) or activation markers such as neopterin. These can be used as early biomarkers of infection, or to evaluate or predict the clinical response to antiviral therapy with interferon-alpha or beta, which are often measured in conjunction with detection of neutralizing antibodies. Tumor necrosis factor alpha (TNFa) and the acute phase protein, C-reactive protein (CRP), are also used as biomarkers of inflammation linked to infective exacerbations. The nanowire arrays will be used to quantify levels of CRP, TNFa, neopterin, IP-10 and beta2-microglobulin (nanowire functionalization with specific antibodies) and levels of antibodies to interferon-beta (functionalization with interferon-beta itself). The project will also explore the potential of DNA aptamers as synthetic antibodies, a more costeffective and nanowire-compatible approach than classical protein antibodies. Many diseases do not have a single biomarker that correlates with disease state, hence the need to use a panel of biomarkers for more accurate diagnosis and management of disease.The project will have unique access to clinical samples -serum and induced sputum- obtained from patients admitted to the Acute Medical Unit at Southampton General Hospital suffering from acute asthma exacerbations, 60-80% of which are triggered by a respiratory virus, and following their recovery. We will also have access to serum samples from asthmaticvolunteers undergoing Phase I clinical trials using inhaled interferon-beta which is being developed for treatment of virusinduced asthma exacerbations. Results will be directly compared against data obtained using traditional methods (ELISA) and to clinical outcomes (e.g. respiratory virus detection, exacerbation severity, response to treatment).