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，全球性传播感染性传播感染的患病率在英国显着增长，例如梅毒、衣原体和淋病分别增加了 1800%、150% 和 42%，导致全国泌尿生殖医学 (GUM) 诊所进行了超过 200 万次检测。能够在护理点使用快速、经济有效且稳健的检测方法来检测所有这些和其他性传播感染，不仅可以加强患者护理服务的提供，而且还有助于减少感染的进一步传播。目前不存在这样的测试，当前的方法最多只能提供双重目标感染测定，但无法满足基于实验室的测试的灵敏度和特异性。因此，面临的挑战是开发具有成本效益、信息丰富的自动化测试，这些测试与实验室中进行的测试一样准确，并且可以在初级和二级护理环境中使用。因此，该项目的目标是通过整合两个现有的互补流程（Hull 的 DNA 提取和 PCR 扩增芯片以及 Randox 的 DNA 杂交阵列和基于化学发光的检测器）来实现这一未满足的挑战，以创建适当的技术。 Randox - Hull 联盟不仅拥有开发此类技术的技术和工程能力，而且还通过 Randox 在该领域的商业存在开辟了清晰的市场路线。基于最近资助的研究（EPSRC EP/H007385/1、EP/D040930 和 BBSRC BBE0027221），该项目将开发单次诊断芯片，该芯片将预载所有必要的试剂和废物遏制要求，以便与专门设计的便携式低功耗控制和数据处理仪器结合使用。实际上，用户只需将尿液或拭子样本直接添加到芯片上样本引入室中的吸附剂中。这种水平的样品处理和片上处理是创造技术的关键，该技术实际上可以减少在实验室环境外使用时潜在的感染风险和样品之间的交叉污染。手动关闭样品室将通过与预装的盐酸胍混合来启动样品中 DNA 的释放，DNA 被捕获在二氧化硅整料上，从那里它将被洗脱到含有预装试剂的 PCR 室中。扩增后，生物素标记的 DNA 靶标将通过电泳输送到检测室，以避免体积变化，杂交反应将它们与一系列特定的互补核苷酸序列连接。然后，控制箱中的执行器将通过流体动力输送芯片上的试剂和清洗溶液，以生成化学发光信号，用于阵列上存在的对照和目标分析物的任何阳性杂交，并使用 CCD 相机进行检测。该芯片将安装在控制仪器中，该仪器将容纳所需的电源、压力执行器、电阻加热风冷热循环仪和 CCD 检测器。将开发软件来控制所提议仪器的测序和数据采集，以及合适的用户界面，以便在临床医生看病人之前向他们提供信息。