STTR PHASE I: Nano-Needle DNA Biosensor For Iin-Situ Direct Detection

该 STTR 第一阶段研究计划将证明纳米级针（纳米针）探针生物传感器在不放大目标/信号的情况下测定低丰度核酸的可行性。该工作的独特之处在于结合使用介电泳和毛细管作用来制造由混合纳米材料制成的高纵横比纳米针，并将纳米针用作生物传感器，通过尺寸排他性的样品浓缩来实现高灵敏度。该公司已经成功地对以定量方式掺入缓冲液或细胞溶液中的经嵌入剂处理的 lambda-DNA 进行非特异性采样/分析。这一成果的灵敏度约为 10pg/mL (0.3fM)，与天然存在的 DNA 种类的浓度相当，足以筛选/检测血液中的循环 DNA（低于 50ng/mL）。 该方法将为高纵横比纳米针结构的高通量制造及其在生物传感平台上的应用铺平道路。所提出的纳米针生物传感器装置将实现简单、快速而灵敏的检测，无需目标/信号放大，并最大限度地减少作为现场可部署装置的物理尺寸、相应的能耗、样品尺寸和样品制备时间的操作。该生物传感器旨在使用经过最低限度处理的样品或原始样品进行即时检测。最终，该设备将适用于核酸检测（NAT），以通过最少处理的样本来快速筛查疾病（例如癌症）。

This STTR Phase I research proposal will demonstrate the feasibility of a nanoscale needle (nano-needle) probe biosensor assaying low-abundance nucleic acids without target-/signal amplification. The unique point in the proposed work lies in the combined use of dielectrophoresis and capillary action to fabricate a high-aspect ratio nano-needle made of a hybrid nanomaterial and to operate the nano-needle as a biosensor, achieving high sensitivity through size-exclusive sample concentration. The company has already succeeded in nonspecifically sampling/assaying intercalator-treated lambda-DNA spiked into a buffer or cell solution in a quantitative manner. The sensitivity of this achievement was around 10pg/mL (0.3fM) which is comparable to the concentration of naturally occurring DNA species and sufficient for screening/detection of circulating DNA in blood (sub-50ng/mL). The method will pave the way to high throughput fabrication of high aspect ratio nano-needle structures and the application to the biosensing platform. The proposed nano-needle biosensor device will enable simple, rapid, yet sensitive detection without target-/signal amplification and minimize the operation in terms of the physical size, corresponding energy consumption, sample size, and sample preparation time as a field-deployable device. The biosensor is aimed for point-of-care-testing using minimally treated or raw samples. Eventually, the device will be applicable to nucleic acid testing (NAT) for rapid screening of diseases (e.g. cancer), which is enabled through minimally treated samples.