Sensor Technology for Rapid Microbial Identification and Susceptibility Testing

描述(由申请人提供)： 抗生素耐药病原体的出现是一场全球医疗危机，迫使医生用更有效的抗生素治疗常见的传染病。迫切需要新的战略来快速识别耐药的医疗保健相关感染，并为临床医生提供实时信息来指导抗生素的选择。这项合作提案中描述的研究的目标是开发所有必要的技术组件，以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。我们已经组建了一个多学科团队来实现这一目标，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.)、药敏试验(D.B.，J.H.)、泌尿科(B.C.)和生物数学(E.L.)。该生物传感器小组已成功开发出一种用于快速鉴定泌尿系统病原体基因型别的电化学传感器。电化学传感器分析包括目标16S rRNA与物种特异性捕获和检测器探针的“夹心”杂交。该检测可以在室温下进行，对缺陷的敏感性只有200个细菌。对尿路感染患者的尿液样本的临床研究已经证明了概念的正确性。 《研究计划》有三个具体目标。具体目标1描述了通过改进对表面化学和信号放大技术的控制来提高电化学传感器分析性能的方法。这些努力将导致在不增加背景信号的情况下获得类似于PCR的灵敏度。具体目的2描述了一种快速抗菌药敏感性检测方法的发展。针对RFA中要求的大多数病原体已经开发了物种特异性探针，这些探针将在快速测试临床分离株对相关抗生素的表型反应时进行验证和测试。具体目标3将是使电化学传感器分析适应自动化、机器人、高通量系统，我们称之为PATHOSENSE仪器。PATHOSENSE仪器的分析性能将在医院获得性尿路感染高危患者的临床研究中进行评估。正如产品开发计划中所述，这些研究将使我们的工业合作伙伴GeneFluidics能够在多种临床环境中近期部署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.