SBIR Phase II: Biosensor for Label-Free, Real-Time Monitoring of Environmental Pathogens

这个小型企业创新研究第二阶段项目将开发一种便携式系统，用于现场实时、同时检测和识别水或气溶胶样本中的多种环境微生物和毒素，具有高灵敏度和特异度，并将假阳性或阴性事件降至最低。该系统由一次性生物传感器芯片和光学阅读器组成。这种探测是基于一种被称为光栅耦合表面等离子体共振成像(GCSPRI)的专有光学转换技术。先前的第一阶段工作已经证明了GCSPR微阵列技术用于高灵敏度多路检测的可行性。第二阶段项目的目标是开发检测/识别传感器的实验室原型和原型芯片，用于对包括细菌、病毒和毒素在内的三种分析物的模型集进行多重检测。非致病生物将被用作模型系统。将探索多种表位检测方法，以减少误报的可能性。第二阶段工作的最终结果将是使用手动样品引入的实验室原型进行演示。这将为便携式检测系统的商业化开发提供一个合乎逻辑的关键里程碑，该系统与气溶胶收集器接口，用于现场测试和评估。该项目的商业应用是国土防务的生物制剂检测。当需要对污染环境进行快速评估时，近乎实时的、具有低误警率的多路测量的能力将是很有价值的。潜在的应用将包括医院，在那里可能会出现医院感染；大型建筑物，在那里意外污染霉菌孢子、军团菌和其他病原体可能会对健康造成危害；娱乐用水和饮用水供应，其中水传播病原体是一个非常令人担忧的问题；以及食品业，在那里需要灵敏的方法来在线和实时检测病原体。

This Small Business Innovation Research Phase II project will develop a portable system for real-time, simultaneous detection and identification of multiple environmental microbes and toxins from aqueous or aerosol samples, on site, with high sensitivity and specificity and with minimal false positives or negative events. The system consists of a disposable biosensor chip and an optical reader device. The detection is based on a proprietary optical transduction technology known as grating-coupled surface plasmon resonance imaging (GCSPRI). Prior Phase I work has demonstrated the feasibility of the GCSPR microarray technology for multiplexed detection with high sensitivity. The goal of the Phase II project is to develop a laboratory prototype of a detection/identification sensor and a prototype chip for multiplexed detection of a model set of three analytes including a bacterium, a virus and a toxin. Non-pathogenic organisms will be used as model systems. Multi-epitope detection methods will be explored for reducing the probability of false alarms. The end result of the Phase II effort will be a demonstration with the laboratory prototype using manual sample introduction. This will provide the logical and critical milestone to transition into commercial development of a portable detection system interfaced to an aerosol collector for field testing and evaluation.The commercial application of this project is in the detection of biological agents for Homeland Defense. The capability for near real-time, multiplexed measurements with a low false alarm rate will be valuable whenever rapid assessment of a contaminated environment is needed. The potential applications would include hospitals, where nosocomial infections may arise; large buildings, where accidental contamination with mold spores, Legionella and other pathogens may create health hazards; recreational water and drinking water supplies, where waterborne pathogens are a great concern; and the food industry, where there is a need for sensitive methods for on-line and real-time detection of pathogens.