Development of diagnostic teststrips incorporating concatameric aptamers for quantitative detection of C. difficile

对传统治疗具有耐药性的新致病物种和菌株的出现是我们这个时代最紧迫的健康问题之一。去年，禽流感病毒的流行和艰难梭菌在加拿大几家医院的爆发是最新的例子，说明了这一点。这些疫情可能造成巨大的经济负担，更重要的是，对人类健康构成巨大风险。早期和准确发现和识别危险病原体是预防大规模疫情的关键步骤。这在今天尤为重要，因为商业全球化和旅行时间的缩短大大增加了传染病传播的速度和广度。使用分子方法（PCR检测DNA）或免疫学方法的传统微生物病原体检测技术需要较高的病原体浓度和/或较长的分析时间——在评估病情恶化的患者或试图确定和控制大规模疫情时，这些时间可能非常宝贵。此外，目前的方法往往不能区分疾病携带者和感染者（艰难梭菌的一个特殊问题）。因此，仍然迫切需要开发快速、灵敏、准确和廉价的检测病原体的方法和设备。同样重要的是，需要一种能够迅速到位的可靠方法，以创建新的传感器和分析方法，以应对未预料到的疫情。这个CRD项目的目标是开发一个基于核酸适体和酶的平台，该平台可以提供核酸和病原体之间选择性结合事件的高扩增，以便在几分钟内在简单的纸试纸上检测到极低水平的病原体。该提案汇集了两名共同申请人，他们在DNA适体和DNA酶技术、生物分子固定化和生物传感器以及生物活性纸质诊断开发方面具有专业知识。该团队将开发和验证新的适体和适体酶传感平台，用于检测病原体，这些平台可以获得加拿大合作伙伴的许可，最终实现商业化。

The emergence of new pathogenic species and strains that are resistant to traditional treatments are amongst the most pressing health concerns of our time. The bird flu virus epidemic and the outbreak of Clostridium difficile in several hospitals across Canada in the past year are the latest examples to drive this point home. These outbreaks can be a huge economic burden and, more importantly, pose a tremendous risk to human health. Early and accurate detection and identification of dangerous pathogens is a crucial step in preventing large-scale outbreaks. This is particularly relevant today as globalization of commerce and decreased travel times have significantly increased the rate and breadth of the spread of infectious agents. Traditional microbial pathogen detection techniques using molecular methods (DNA detection with PCR) or immunological methods require large pathogen concentrations and/or long analysis times - time that can be very precious when assessing a patient with a deteriorating condition or when trying to identify and control a large-scale outbreak. In addition, current methods often cannot distinguish between carriers of a disease and those who are infective (a particular issue with C. difficile). Hence there is still a significant need to develop rapid, sensitive, accurate and inexpensive methods and devices for the detection of pathogens. Equally important is the need for a reliable method that can be put in place quickly to create new sensors and assays to respond to unanticipated outbreaks. The goal of this CRD project is to develop a platform based on nucleic acid aptamers and enzymes that can provide high amplification of a selective binding event between the nucleic acid and the pathogen to allow detection of very low levels of pathogens in minutes on a simple paper-based test strip. The proposal brings together two co-applicants with expertise in DNA aptamer and DNA enzyme technology, biomolecule immobilization and biosensors, and development of bioactive paper-based diagnostics. This team will develop and validate new aptamer and aptazyme sensing platforms for detection of pathogens that can be licensed by our Canadian partner for eventual commercialization.