Development of a rapid multiplex Lab on a Chip system for detection of 10 STI pathogens using Biochip Array Technology

开发快速多重芯片实验室系统，利用生物芯片阵列技术检测 10 种 STI 病原体

• 批准号: TS/I00114X/1
• 负责人: Steve Haswell
• 金额: $ 86.95万
• 依托单位: University of Hull
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2010
• 资助国家: 英国
• 起止时间: 2010 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=TS%2FI00114X%2F1
• 关键词: Development; rapid; multiplex; Lab; Chip

Over the past decade the world has seen a significant growth in the prevalence of sexually transmitted infections STIs in the UK for example Syphilis, Chlamydia and Gonorrhoea have shown increases of 1800%, 150% and 42%, respectively, resulting in over 2 million tests being performed in genito-urinary medicine (GUM) clinics across the nation. The availability of a rapid, cost-effective and robust test for the detection of all of these and other STIs, which could be used at the point of care, would not only enhance the delivery of patient care services but would also aid in reducing the further spread of infection. At present no such test exists, with current methodology offering at best duplex target infection assays that fail to meet the sensitivity and specificity of laboratory based tests. The challenge is therefore to develop cost-effective, automated information rich tests that are as accurate as those conducted in the laboratory and that can be used in both primary and secondary care settings. The aim of this project is therefore to realise this unmet challenge through the integration of two existing and complimentary processes (the DNA extraction and PCR amplification chip from Hull and the DNA hybridisation array and chemiluminescence based detector from Randox), to create appropriate technology. The Randox - Hull consortium not only has the technical and engineering capability to develop such technology but also through Randox's commercial presence in the field a clear route to market. Based on recently funded research (EPSRC EP/H007385/1, EP/D040930 and BBSRC BBE0027221), this project will develop a single shot diagnostic chip, which will come pre-loaded with all the necessary reagents and waste containment requirements for it to operate in conjunction with a specifically designed portable low power control and data processing instrument. In practice the user will only need to add a urine or swab sample directly to an adsorbent in a sample introduction chamber on the chip. This level of sample handling and on-chip processing is key in creating technology that will practically reduce both potential exposure to infection and cross contamination between samples when used out of the laboratory environment. Manual closure of the sample chamber will initiate the release of DNA from the sample by mixing with pre-loaded guanidine hydrochloride with the DNA being trapped on a silica monolith from where it will be eluted into a PCR chamber containing pre-loaded reagents. Following amplification the biotin-labelled DNA targets will be transported electrophoretically, to avoid volumetric changes, into the detection chamber where a hybridisation reaction will tether them to an array of specific complimentary nucleotide sequences. Actuators in the control box will then hydrodynamically deliver reagent and wash solutions held on chip to generate a chemiluminescence signal for any positive hybridisation of controls and target analytes present on the array which will be detected using a CCD camera. The chip will be located in a control instrument which will house the required power supplies, pressure actuators, resistively heated air cooled thermocycler and the CCD detector. Software will be developed to control the sequencing and data acquisition of the proposed instrument together with a suitable user interface for presenting information to a clinician prior to them seeing a patient.

在过去的十年中，世界在英国性传播感染性传播感染的流行率显着增长，例如，梅毒，衣原体和淋病的疾病分别增加了1800％，150％和42％的增长，导致在整个国家（Gum）中进行了超过200万种测试。可以在医疗点使用所有这些和其他性传播感染的检测快速，成本效益且可靠的测试，不仅可以增强患者护理服务的提供，而且还将有助于减少感染的进一步传播。目前尚无这样的测试，目前的方法学提供了最佳双链目标感染测定法，但无法满足基于实验室测试的敏感性和特异性。因此，面临的挑战是开发具有成本效益的自动化信息丰富的测试，这些测试与实验室中的测试一样准确，并且可以在初级和二级护理环境中使用。因此，该项目的目的是通过整合两个现有的和免费的过程（来自Hull的DNA提取和PCR扩增芯片，DNA杂交阵列和基于Randox的化学发光的检测器）来实现这一未满足的挑战，以创建适当的技术。 Randox-赫尔财团不仅具有开发此类技术的技术和工程能力，而且还具有Randox在该领域的商业形象，这是一条清晰的市场途径。基于最近资助的研究（EPSRC EP/H007385/1，EP/D040930和BBSRC BBE0027221），该项目将开发单个Shot诊断芯片，该芯片将预装，该芯片将预加载所有必要的试剂和浪费遏制要求，以便与特定设计的低功能处理能力控制和数据处理量和数据处理。实际上，用户只需要将尿液或拭子样品直接添加到芯片上样本简介室中的吸附剂中。这种样品处理和片上处理是创造技术的关键，该技术实际上将减少在实验室环境中使用时样品之间潜在的感染和交叉污染。样品室的手动闭合将通过与预加载的盐酸盐酸盐盐混合DNA与DNA被困在二氧化硅整体上，从而从样品中释放DNA，从而将其从中洗脱成含有预载试剂的PCR室。放大后，生物素标记的DNA靶标将通过电泳运输，以避免体积变化，进入检测室，其中杂交反应将它们束缚在一系列特定的免费核苷酸序列中。然后，控制盒中的致动器将流体动力的试剂和洗涤溶液固定在芯片上，以产生化学发光信号，以用于对照组的任何正杂交和阵列上存在的目标分析物的正杂交，并使用CCD摄像头检测到。芯片将位于控制仪器中，该仪器将容纳所需的电源，压力执行器，电阻加热的空气冷却热环体和CCD检测器。将开发软件，以控制所提出的仪器的测序和数据采集以及合适的用户界面，以在看到患者之前向临床医生提供信息。