Uropathogen Detection Using DNA Biosensors

描述(申请人提供)：尿路感染最多 美国常见的泌尿系疾病，是患者的主要原因 发病率和医疗保健支出。这个生物工程研究伙伴关系 提案涉及开发一种微型机电系统，用于 尿路病原菌的基因分型检测和种属特异性鉴定 时间范围(从样本采集到读数5-10分钟) 护理点诊断和治疗。这项提议的重点是生产 一种自给式微生物病原体检测装置及其检测方法 使用临床尿液样本进行检测。加州大学洛杉矶分校的研究提供了两个关键 使尿路病原体传感器的开发成为可能的技术进步。 首先是用于样品处理的微流控技术。第二个是一个 电化学微传感器，允许超灵敏地检测特定的 DNA-RNA或DNA-DNA杂交事件，不需要靶标 放大。这个项目已经开发了一年多，涉及一个 多学科努力，包括微流体和微流体领域的领导者 微传感器技术、分子微生物学、儿科学和生物数学。 具体目标1描述微流体学研究将如何应用于 用于尿路病原菌浓缩、微量混合的错流过滤器的研制 用于尿路病原体核酸的处理，以及传感器表面的清洗。 具体目标2涉及微传感器阵列的制造、开发 链霉亲和素自组装单分子膜及寡核苷酸探针检测 用于微传感器上尿路病原体rRNA和mRNA的电化学检测 浮出水面。具体目标3将涉及微流体和 传感器组件及其在模拟和实际上的分析有效性检验 尿液样本。具体目标4将涉及批量制造该装置 以及临床检查尿传感器结果与 尿路感染的临床相关因素。

DESCRIPTION (provided by the applicant): Urinary tract infection is the most common urological disease in the United States and is a major cause of patient morbidity and health-care expenditure. This Bioengineering Research Partnership proposal involves development of a microelectromechanical system for the genotypic detection and species-specific identification of uropathogens within a time frame (5-10 minutes from sample collection to readout) that would enable point-of-care diagnosis and treatment. The focus of this proposal is to produce a self-contained microbial pathogen detection device and to examine its performance using clinical urine samples. Research at UCLA has provided two key technological advances that make development of a uropathogen sensor feasible. The first is microfluidics for sample processing. The second is an electrochemical microsensor which allows ultrasensitive detection of specific DNA-RNA or DNA-DNA hybridization events, without the need for target amplification. This project has been in development for over a year involving a multidisciplinary effort including leaders in the fields of microfluidics and microsensor technology, molecular microbiology, pediatrics and biomathematics. Specific Aim 1 describes how microfluidics studies will be applied to development of a crossflow filter for uropathogen concentration, micromixing for processing of uropathogen nucleic acids, and washing of the sensor surface. Specific Aim 2 involves fabrication of the microsensor array, development of a streptavidin self-assembled monolayer, and testing of oligonucleotide probes for electrochemical detection of uropathogen rRNA and mRNA on the microsensor surface. Specific Aim 3 will involve integration of the microfluidics and sensor components and testing of its analytic validity on simulated and actual urine specimens. Specific Aim 4 will involve batch fabrication of the device and clinical examination of the association between urosensor results and clinical correlates of urinary tract infection.