Development of a polymer-based sensing platform for the thermal detection of antimicrobial resistance

抗生素彻底改变了现代医学，但这些“神奇”药物正受到威胁，因为抗生素耐药菌株的迅速出现不再对标准抗生素治疗产生反应。这危及目前的标准程序，如大手术，癌症治疗和器官移植。监测这些耐药菌株是对抗它们的关键。在这项提案中，我们将生产一种生物传感器，用于检测细菌，特别是那些具有抗菌素耐药性的细菌。我们可以以简单和低成本的方式快速确定细菌感染的来源，使临床医生能够制定个性化的治疗方案，从而使患者的护理受益。此外，我们将把它扩展到一个阵列格式，用于同时检测细菌和抗生素，这可以用于筛选（食品）样品中的抗生素残留，并将提供有价值的洞察细菌如何发展AMR特性。我们将使用一种称为分子印迹的技术来生产传感器平台。这些分子印迹聚合物（MIP）通常被称为“塑料”抗体。这些材料具有多孔结构，具有针对其靶分子的高亲和力结合位点。与“天然”抗体相比，它们的优势包括低成本、简单的制备、鲁棒性和在极端环境（pH、不利温度和有机溶剂）中工作的能力。PI小组先前的工作表明，目标与印迹聚合物的结合可以改变通过聚合物的热传导，基本上阻止热流。这会导致温差，可通过热传感器（热电偶装置）测量。热流的这种变化取决于目标浓度。这种方法，专利的热传递方法（HTM），只研究了MIP微结构。在这项提案中，我们将采取一种新的电化学方法来开发MIP纳米层，这将增加所开发的传感器平台的灵敏度。该项目包括以下步骤：（a）使用电化学方法制备MIP传感器。我们将从文献数据库中确定的五种不同单体制备纳米厚的细菌印迹层功能化到电极上，以结合细菌。使用HTM，将确定哪种单体具有最高的结合特定细菌菌株的潜力，从而使我们能够优化MIP。将测量一系列与医学相关的目标（包括金黄色葡萄球菌菌株，其中一些菌株表现出抗菌素耐药性），并在时间，选择性和亲和力方面优化传感器性能。(b)缓冲溶液中细菌的热测量我们将使用MIP传感器（六种细菌库）进行热测量，以评估缓冲溶液中的细菌负荷。这些测量将根据当前的金标准技术（ELISA，基因分型）进行验证，以确定开发的热传感策略的准确度和精密度。(c)“复杂”样品的热测量临床或食品样品是复杂的基质-我们将评估是否可以在存在过量其他（无害）细菌的情况下选择性地检测某些细菌菌株。最后，我们将探索是否可以通过整合抗生素化合物的特异性MIP，将这种传感器转化为同时检测细菌和抗生素的阵列形式。这一提议将建立PI的研究组合，建立她的独立性，并为多学科和令人兴奋的研究计划奠定基础。马斯特里赫特的一个项目合作伙伴将提供热测量方面的建议，并为知识交流访问提供服务。开发的传感器平台具有商业潜力，因为其成本低且简单，PI将在项目时间轴期间对其进行探索。

Antibiotics revolutionized modern medicine, but these 'wonder' drugs are under threat due to the rapid emergence of antimicrobial resistant bacterial strains that no longer respond to standard antibiotic treatment. This endangers current standard procedures, such as major surgery, cancer therapy and organ transplantation. Monitoring these resistant strains is key to combating them.In this proposal, we will produce a biosensor for the detection of bacteria, particularly those with antimicrobial resistance. In a simple and low-cost manner, we can rapidly identify the source of bacterial infection to enable clinicians to develop a personalized treatment plan that will benefit patients' care. In addition, we will expand this to an array format for the simultaneous detection of bacteria and antibiotics, which can serve to screen (food) samples for antibiotic residues and will provide valuable insight into how bacteria develop AMR properties. We will use a technique called molecular imprinting for producing the sensor platform. These Molecularly Imprinted Polymers (MIPs) are often referred to as "plastic" antibodies. These materials have a porous structure, with high affinity binding sites for their target molecule. Their advantages over "natural" antibodies include low-cost, straightforward preparation, robustness, and ability to work in extreme environments (pH, adverse temperatures and organic solvents). Prior work in the PI's group has shown that binding of targets to imprinted polymers can alter the conduction of heat through the polymer essentially blocking heat-flow. This can lead to a temperature differential which can be measured by a thermal sensor (thermocouple device). This change in heat-flow is dependent on target concentration. This method, patented as the Heat-Transfer Method (HTM), has only been studied with MIP microstructures. In this proposal, we will take a novel electrochemical approach to develop MIP nanolayers that will increase the sensitivity of the developed sensor platform. This project consists of the following steps: (a) Use of electrochemical methods to prepare MIP sensors.We will prepare nanometre thick bacterial imprinted layers functionalised onto electrodes from five different monomers, which have been identified from literature databases to bind bacteria. Using HTM it will be determined which monomer has the highest potential to bind a particular bacterial strain allowing us to optimise the MIP. A series of medically relevant targets (including Staphylococcus aureus strains, some of which exhibit antimicrobial resistance) will be measured and the sensor performance will be optimised in terms of time, selectivity and affinity.(b) Thermal measurements of bacterial in buffered solutionsWe will perform thermal measurements with the MIP sensors (library of six bacteria) to evaluate the bacterial loads in buffered solutions. These measurements will be validated against current gold-standard techniques (ELISA, genotyping) to determine the accuracy and precision of the developed thermal sensing strategy. (c) Thermal measurements of "complex" samplesClinical or food samples are complex matrices - we will evaluate if we can selectively detect certain bacterial strains in the presence of an excess of other (harmless) bacteria. Finally, we will explore if we can transform this sensor into an array format for the simultaneous detection of bacteria and antibiotics, by integrating MIPs specific for antibiotic compounds.This proposal will build the research portfolio of the PI, establish her independence, and lay the foundation of a multidisciplinary and exciting research programme. A project partner at Maastricht will provide advice on thermal measurements and serve for knowledge exchange visits. The developed sensor platform has commercial potential due to its low-cost and simplicity and the PI will explore its this during the project timeline.