SBIR Phase I :Point-of-Care Test for Identifying Gram-Negative Urinary Tract Infections in Companion Animals

这个SBIR一期项目将开发一种手持式、即时护理、传感条和仪器，用于立即识别引起伴侣动物尿路感染（UTI）的细菌种类。这项技术将允许兽医在初步检查时明智地为动物开抗生素。仅在美国，每年就有近150万名兽医因狗和猫的尿路感染而就诊。获得最初尿路感染检测结果的平均时间为48小时，在此期间动物通常服用抗生素。不到一半的尿液培养显示有细菌生长，这表明大多数抗生素处方是不必要的。此外，确定引起尿路感染的细菌种类是很重要的，这样就可以使用适当的抗生素。谨慎使用抗生素将控制感染的传播和抗生素耐药性的出现，从而使动物更健康，并减少对新抗生素药物开发的需求。这个项目将帮助翻译最初的研究和发现，这些研究和发现以前是由国家科学基金会资助的。这项技术的基本原理是，每一种细菌都产生和排泄自己独特的一套化学物质，用于协调细胞间的活动，类似于动物产生的信息素。通过检测这些化学物质，可以确定样品中存在哪些细菌细胞。样品中的化学物质比细菌细胞的数量要多得多，这使得这种方法比其他技术更敏感，更容易实施。这项研究将建立一个无标签的、基于适体体的电化学传感平台，用于在收集样本的两分钟内检测和鉴定引起伴侣动物尿路感染的四种最常见的革兰氏阴性细菌病原体。该方法的新颖之处在于测量细菌分泌的独特群体感应分子（QSMs）。这些qsm由所有细菌细胞以大约每小时10,000分子的速度连续产生，从而将检测极限提高了四个数量级。一组电极，用新型适配体修饰，用电活性分子功能化，将用于检测每一种细菌产生的qsm，作为生物标志物。将优化的适体分子与电化学传感和专有的信号处理算法相结合，可以实现前所未有的检测，灵敏度和特异性限制。该技术将直接在复杂流体中工作，无需添加任何试剂或分离步骤。

This SBIR Phase I project will develop a hand-held, point-of-care, sensing strip and instrument for immediate identification of which bacterial species are causing a urinary tract infection (UTI) in a companion animal. This technology will allow veterinarians to judiciously prescribe antibiotics for an animal during the initial examination. In the United States alone, there are nearly 1.5 million veterinary visits annually for UTIs in dogs and cats. The average time to initial UTI test results is 48 hours, during which time the animal is generally consuming antibiotics. Less than half of urine cultures show any bacterial growth, indicating that the majority of antibiotic prescriptions were unnecessary. Further, it is important to identify which bacterial species are causing the UTI, so that the proper antibiotic can be administered. Prudent use of antibiotics will control the spread of infection and the emergence of antibiotic resistance, thus resulting in healthier animals and reducing the need for new antibiotic drug development. This project will help translate the initial research and discoveries that were funded previously by the National Science Foundation. The underlying principle for this technology is that each bacterial species produces and excretes its own unique set of chemicals that are used to coordinate intercellular activities, analogous to pheromones produced by animals. It is possible to identify what bacterial cells are present in a sample by detecting these chemicals. There are much greater quantities of the chemicals in the sample than bacterial cells, making this approach more sensitive and easier to implement than other technologies.The proposed research will result in the creation of a label-free, aptamer-based, electrochemical sensing platform for the detection and identification of the four most common Gram-negative bacterial pathogens causing UTIs in companion animals, within two minutes of sample collection. The novelty of the approach is to measure unique quorum sensing molecules (QSMs) that are secreted by the bacteria. These QSMs are produced by all bacterial cells continuously at a rate of approximately 10,000 molecules per hour, thereby improving the limit of detection by four orders of magnitude. An array of electrodes, modified with novel aptamers and functionalized with electro-active molecules, will be used to detect the QSMs, acting as biomarkers, produced by each bacterial species. Combining optimized aptamer molecules with electrochemical sensing and a proprietary signal processing algorithm allows for unprecedented limits of detection, sensitivity, and specificity. The technology will work directly in complex fluids without any reagent addition or separation steps.