An Integrated Diagnostic Biochip for Point of Care Pathogen Identification

描述（由申请人提供）：迫切需要临床有用的护理点（POC）技术来鉴定病原体，特别是在非传统医疗保健环境中。本合作研究计划的目标是开发一个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），它是一个完善的、经过临床验证的电化学生物传感器阵列，集成了微流体技术，结合了利用电动力加速样品处理的创新策略。我们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（M.S.）方面建立了学术和行业合作伙伴关系。我们的团队先前描述了用于快速识别临床尿液样本中的尿路病原体的电化学生物传感器。该实验包括将细菌目标16S rRNA与物种特异性捕获和检测器探针进行夹心杂交。目前，由于需要手动样品处理步骤，包括离心、混合和试剂转移，该检测方法不适用于POC应用。需要额外的技术开发和系统集成来提高灵敏度、速度和实现POC效用。我们提出了四个具体目标：(1)使用多功能“通用电极”开发电动和微流体样品处理模块；(2)研制了由通用电极阵列、样品处理流体处理盒和试剂存储盒组成的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)使用加标尿液样本和未知临床尿液样本验证P-PAL。我们预计将在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的密切合作，我们为未来的临床部署提出了一个基本的产品开发计划。

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.