An Integrated Diagnostic Biochip for Point of Care Pathogen Identification

描述（由申请人提供）：迫切需要用于病原体识别的临床有用的床旁（POC）技术，特别是在非传统医疗环境中。这项合作研究提案的目标是开发一种高度便携的诊断平台，用于尿路感染（UTI）的POC诊断。该平台的核心是便携式病原体警报实验室（P-PAL），它是一种成熟的、经临床验证的电化学生物传感器阵列，与微流体技术相结合，并结合创新策略，利用动电力加快样品处理。我们已经组建了一个学术界与工业界的合作伙伴关系，在电化学生物传感器（V.G.），微流体/电动力学（P.W.），泌尿学（J.L.），临床微生物学（E.J.B.），和生物统计学（硕士）。我们的团队先前描述了用于快速鉴定临床尿液样本中的尿路病原体的电化学生物传感器。该测定涉及细菌靶16 S rRNA与种特异性捕获和检测探针的夹心杂交。目前，鉴于需要手动样品处理步骤，包括离心、混合和试剂转移，该测定法不适用于POC应用。需要额外的技术开发和系统集成，以提高灵敏度，速度和启用POC实用程序。我们提出了4个具体目标：（1）开发使用多功能“通用电极”的电动和微流控样品处理模块：（2）开发由通用电极阵列、样品处理流体处理盒和试剂储存盒组成的P-PAL盒;（3）整合P-PAL系统的所有流体、气动和电气控制，用于试剂输送、试剂-样品混合和目标检测;（4）用加标尿样和未知临床尿样验证P-PAL。我们预计，我们将在15分钟内实现分子病原体鉴定，检测灵敏度为1000 cfu/ml。通过与我们的行业合作伙伴GeneFluidics密切合作，我们提出了未来临床部署的基本原理产品开发计划。

DESCRIPTION (provided by applicant): There is an urgent need for clinically useful point of care (POC) technologies for pathogen identification, particularly in nontraditional healthcare settings. The goal of this collaborative research proposal is to develop a highly portable diagnostic platform for POC diagnosis of urinary tract infections (UTI). The core of this platform, Portable Pathogen Alert Laboratory (P-PAL), is a well-established, clinically validated electrochemical biosensor array integrated with microfluidic technology combined with innovative strategies to expedite sample processing using electrokinetic forces. We have assembled an academic-industry partnership with complementary expertise in electrochemical biosensors (V.G.), microfluidics/electrokinetics (P.W.), urology (J.L.), clinical microbiology (E.J.B.), and biostatistics (M.S.). Our team previously described the electrochemical biosensor for rapid identification of uropathogens in clinical urine samples. The assay involves sandwich hybridization of bacterial target 16S rRNA to species-specific capture and detector probes. At present, the assay is not practical for POC application given the requirement for manual sampleprocessing steps including centrifugation, mixing and reagent transferring. Additional technology development and system integration are needed to enhance the sensitivity, speed and to enable POC utility. We propose 4 Specific Aims: (1) To develop electrokinetic and microfluidic sample processing modules using a multi-functional 'universal electrode'; (2) To develop the P-PAL Cartridge composed of the universal electrode array, the sample processing fluidic handling cartridge and a reagent storage cartridge; (3) To integrate all the fluidic, pneumatic, and electrical controls for the P-PAL System for reagent delivery, reagent-sample mixing, and target detection; and (4) To validate P-PAL using spiked urine samples and unknown clinical urine samples. We anticipate that we will achieve molecular pathogen identification within 15 minutes with detection sensitivity of 1000 cfu/ml. In close collaboration with our industry partner GeneFluidics, we propose a rationale Product Development Plan for future clinical deployment.