RAPID point-of-care infection detection and antibiotic-resistance TESTing enabled with laser-patterned microfluidic devices (RAPID-TEST)

这项建议的目的是使用我们的专利激光打印技术来开发低成本的纸质诊断测试，以满足真正未得到满足的医疗需求，即快速识别细菌感染和抗生素敏感性曲线，以指导治疗决策。我们的医疗点(POC)测试将适合在诊所使用，或由患者自己作为会诊前筛查过程的一部分或持续的家庭监测使用。我们已经开发了我们的纸张测试平台，并展示了出色的微流体特性，建立了设备可以渗透到能够检测炎症标志物C反应蛋白的比色系统的原理证明。我们现在正在这个概念的基础上开发令人兴奋的雄心勃勃和概念新颖的诊断测试，供POC使用。我们的目标疾病尿路感染(UTIs)和囊性纤维化(CF)中的慢性呼吸道疾病本身就是严重的，目前处理不佳，但重要的是，成功的技术也将适用于大量其他医疗和工业应用。细菌感染影响了大量的人，带来了重大的生活质量和医疗成本后果。在过去的1-20年里，几乎没有开发出新的抗生素，而且人们越来越关注抗菌素耐药性(AMR)的全球流行。这一问题的一个主要部分直接与广谱、非靶向抗生素的广泛和不分青红皂白地使用有关。然而，在缺乏准确诊断的情况下，选择正确的抗生素是困难的。目前用于鉴定感染病原体和后续测试其对抗生素敏感性的方案是耗时的，并且需要基于微生物培养的专家程序。这些方法不仅昂贵和不方便，而且有一段时间的诊断不确定性，在此期间，治疗方案的选择是经验性的，可能是次优的。患者的健康和福祉受到不利影响，这些延迟导致了AMR的出现。我们提出的基于微流体的新型设备将独特地服务于双重目的-首先，快速、POC识别病原体，其次，廉价和快速地检测其抗生素耐药性。我们将通过使用优化的酶联免疫吸附试验(ELISA)来实现这一点，以在与选定的细菌抗原接触时立即产生颜色变化。我们选择了三种引起尿路感染和CF患者严重感染的问题细菌。一旦单个探测系统得到优化，下一个任务将是将它们多路传输到同一设备上。再加上检测系统，将以显色、琼脂为基础的培养井含有各种抗生素。在短时间的培养后，大约24小时，颜色的变化将表明细菌对哪些抗生素具有抗药性。这一过程将大大减少目前3-4天的诊断时间。这些工具对于全科医生/咨询人员来说将是重要的和及时的，可以为他们的患者感染提供准确的抗生素治疗，并显著节省医疗成本。对于患者来说，症状将会减少，从而提高生活质量。在CF这一终生疾病的背景下，患者可以在自己的家中使用这些设备进行长期监测，方式与英国CF Trust的旗舰SmartCare计划非常一致；这不仅将使患者能够自我管理，促进更早的治疗，而且我们最终可能能够允许未感染的患者相互接触--缺乏对个人感染状况的实时了解目前要求严格隔离，这对患者的福祉产生了负面影响。

The aim of this proposal is to use our proprietary laser printing technique to develop low-cost, paper-based diagnostic tests addressing a genuine unmet healthcare need, namely the rapid identification of bacterial infections and antibiotic susceptibility profiles to guide therapeutic decision making. Our point-of-care (POC) tests will be suitable for use in a clinic or by the patients themselves as part of a pre-consultation screening process or on-going home monitoring. We have already developed our paper testing platform and demonstrated excellent microfluidic properties, establishing the proof of principle that the devices can be impregnated with a colorimetric system capable of detecting the inflammatory marker, C-reactive protein. We are now building on this concept for developing excitingly ambitious and conceptually novel diagnostic tests for use at the POC. Our target diseases, urinary tract infections (UTIs) and the chronic airway disease in cystic fibrosis (CF) are in themselves serious and currently sub-optimally managed, but importantly, the successful technology will also be applicable to a large number of other medical and industrial applications. Bacterial infections affect large numbers of people, with significant quality of life and healthcare cost consequences. There have been very few new antibiotic agents developed over the last 1-2 decades and there is increasing concern over the global epidemic of antimicrobial resistance (AMR). A major part of this problem relates directly to the widespread and indiscriminate use of broad-spectrum, non-targeted antibiotics. Choosing the correct antibiotic is however difficult in the absence of an accurate diagnosis. Current protocols for the identification of an infecting pathogen and follow-on testing of its susceptibility to antibiotics are time-consuming and require specialist microbiology culture-based procedures. These approaches are not only costly and inconvenient, but there is a period of diagnostic uncertainty during which treatment is chosen empirically and may be sub-optimal. Patients' health and well-being is adversely impacted and these delays contribute to the emergence of AMR.Our proposed novel microfluidics-based devices will uniquely serve a dual purpose - first, rapid, POC identification of a pathogen and second, cheap and expedited testing for its antibiotic resistance profile. We will achieve this through the use of an optimised enzyme-linked immunosorbent assay (ELISA) to produce an immediate colour change on contact with the chosen bacterial antigen. We have selected three problematic bacteria causing UTIs and serious infection in patients with CF. Once single detection systems have been optimised, the next task will be to multiplex them onto the same device. Coupled with the detection system will be chromogenic, agar-based culture wells containing various antibiotics. After a short period of incubation, ~ 24 hours, colour changes will indicate the antibiotics to which the bacteria are resistant. This process would significantly reduce the current diagnostic time of 3-4 days.These tools will be important and timely for GPs/Consultants in delivering an accurate antibiotic treatment of their patient's infections with significant savings in healthcare costs. For the patients, there will be reduction in symptoms and consequent improvements in quality of life. In the context of CF, a life-long disease, the devices could be used by patients in their own homes for long-term surveillance, in a fashion very aligned with the UK CF Trust's flagship SmartCare programme; this will not only empower patients in self-management, and facilitate earlier treatment, but we may ultimately be able to allow non-infected patients to have contact with each other - lack of real-time knowledge of an individual's infection status currently mandates strict segregation which has a negative impact on patients' well-being.