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

这项STTRI期研究方案将证明纳米针(纳米针)探针生物传感器在不进行靶/信号放大的情况下分析低丰度核酸的可行性。提出的工作的独特之处在于结合使用介电和毛细管作用来制备由杂化纳米材料制成的高纵横比纳米针，并将纳米针作为生物传感器来操作，通过尺寸专有的样品浓缩来获得高灵敏度。该公司已经成功地以定量的方式对插入缓冲液或细胞溶液中的嵌入剂处理的lambda-DNA进行了非特异性采样/检测。这一成果的灵敏度约为10pg/毫升(0.3fM)，与自然产生的DNA物种的浓度相当，足以筛选/检测血液中循环DNA(低于50 ng/毫升)。该方法为高深宽比纳米针结构的高通量制备及其在生物传感平台上的应用奠定了基础。提出的纳米针生物传感器装置将实现简单、快速、灵敏的检测，而不需要目标/信号放大，并将作为现场可部署装置的物理尺寸、相应的能量消耗、样品大小和样品制备时间的操作最小化。该生物传感器的目标是使用最低限度的处理或原始样本进行护理点测试。最终，该设备将适用于核酸测试(NAT)，用于快速筛查疾病(例如癌症)，这是通过最低限度处理的样本实现的。

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.