Sensor Technology for Rapid Microbial Identification and Susceptibility Testing

描述（由申请人提供）： 抗生素耐药性病原体的出现是一个全球性的医疗危机，迫使医生用更有效的抗生素治疗常见的感染性疾病。迫切需要新的策略来快速识别耐药医疗保健相关感染，并为临床医生提供实时信息以指导抗生素的选择。本合作提案中描述的研究目标是开发生产用于快速高通量细菌病原体鉴定和抗生素敏感性测定的生物传感器所需的所有基本技术组件。为了实现这一目标，我们组建了一个多学科团队，其中包括电化学传感器领域的学术和行业领导者（J.W.，（V.G.）分子微生物学（D. H.），抗微生物敏感性测试（D.B，J.H.），泌尿学（不列颠哥伦比亚省）和生物数学（E. L.）。生物传感器小组成功地开发了一种电化学传感器，用于快速鉴定尿路病原体的基因型。电化学传感器测定涉及靶16S rRNA与物种特异性捕获和检测探针的“夹心”杂交。该测定可在室温下进行，并且对少至200个细菌的缺陷具有灵敏度。概念验证已经在尿路感染患者尿液样本的临床研究中得到证实。 研究计划有三个具体目标。具体目标1描述了通过改进表面化学和信号放大技术的控制来增强电化学传感器测定的性能的方法。这些努力将在不增加背景信号的情况下产生类似PCR的灵敏度。具体目标2描述了快速抗菌药物敏感性测定的开发。已针对RFA中要求的大多数病原体开发了种属特异性探针，这些探针将在快速检测中进行验证和检测，以测量临床分离株对相关抗生素的表型反应。具体目标3将是使电化学传感器测定适应自动化、机器人、高通量系统，我们将其称为PATHOSENSE仪器。将在医院获得性尿路感染高风险患者的临床研究中评价PATHOSENSE仪器的分析性能。如产品开发计划所述，这些研究将使我们的工业合作伙伴GeneFluidics能够在多个临床环境中短期部署PATHOSENSE仪器。

DESCRIPTION (provided by applicant): Emergence of antibiotic resistant pathogens is a global healthcare crisis that is forcing physicians to treat common infectious-diseases with ever more potent antibiotics. New strategies are urgently needed for rapid identification of drug resistant healthcare-associated infections and to provide clinicians with real-time information to guide antibiotic selection. The goal of the research described in this collaborative proposal is to develop all of the essential technological components needed to produce a biosensor for rapid high throughput bacterial pathogen identification and antibiotic susceptibility determination. We have assembled a multidisciplinary team to achieve this goal including academic arid industry leaders in the fields of electrochemical sensors (J. W., V.G.) molecular microbiology (D. H.), antimicrobial susceptibility testing (D.B, J.H.), urology (B.C.) and biomathematics (E. L.). The biosensor group has successfully developed an electrochemical sensor for rapid genotypical identification of uropathogens. The electrochemical sensor assay involves "sandwich" hybridization of target 16S rRNA to species-specific capture' and detector probes. The assay can be performed at room temperature and has the sensitivity to defect as few as 200 bacteria. Proof Of concept has already been demonstrated in a clinical study of urine specimens from patients with urinary tract infections. The Research Plan has three Specific Aims. Specific Aim 1 describes methods to enhance the performance of the electrochemical sensor assay through improved control of the surface chemistry and signal amplification techniques. These efforts will result in PCR-like sensitivity without an increase in background signal. Specific Aim 2 describes development of a rapid antimicrobial susceptibility assay. Species specific probes have been developed for most of the pathogens called for in the RFA, these probes will be validated and tested in a rapid test measuring the phenotypic response of clinical isolates to relevant antibiotics. Specific Aim 3 will be to adapt the electrochemical sensor assay to an automated, robotic, high throughput system, which we refer to as the PATHOSENSE instrument. Analytic performance of the PATHOSENSE instrument will be evaluated in a clinical study of patients at high risk for hospital acquired urinary tract infection. As described in the Product Development Plan, these studies will position our industrial partner, GeneFluidics, for near-term deployment of the PATHOSENSE instrument in multiple clinical settings.