Molecular Beacon Based Extracellular mRNA and Protein Detection for Early Cancer Diagnosis

摘要 包括胰腺癌在内的许多癌症类型具有非常高的死亡率，因为很难检测其 存在于早期阶段。目前胰腺癌的生物标志物，如血清CA-19-9蛋白， 假阳性率和假阴性率。拟议的研究旨在开发一个强大的和用户友好的小说 分子信标和脂质体纳米粒生物芯片技术，并通过检测验证其临床应用潜力 胰腺癌患者血清样本细胞外囊泡中磷脂酰肌醇蛋白聚糖1 mRNA和蛋白的表达 在早期癌症诊断领域的应用迫在眉睫。该项目的具体目标是：目标1： 稳健生物芯片技术的发展。1-1.改进的MB用于高稳定性和错配特异性; 1-2.用于芯片和分析仪校准和分析定量的标准囊泡（SV）; 1-3.非TIRF 荧光显微镜作为一个低成本的分析仪和自动化软件模块，快速和用户友好 图像采集和分析。具体目标2：胰腺癌的多部位技术验证 诊断. 2-1.与qRT-PCR、NGS和ELISA在性能、成本、测定时间和用户方面的比较 友善2-2.多中心盲态验证：OSU的实验室间重复性和盲态验证比较 和使用患者和正常供体血清样品的MSKCC。

Abstract Many cancer types including pancreatic cancer have very high mortality rates because it is difficult to detect their presence in the early stage. Current biomarkers for pancreatic cancer such as serum CA-19-9 protein face high false positive and negative rates. The proposed studies seek to develop a robust and user-friendly novel molecular beacon and lipoplex nanoparticle biochip technology and validate its clinic potential by detecting Glypican 1 mRNA and protein in extracellular vesicles from pancreatic cancer patient serum samples for imminent use in the field of early cancer diagnosis. The specific aims of this project are: Aim 1: Development of Robust Biochip Technology. 1-1. Improved MBs for high stability and mis-match specificity; 1-2. Standard vesicles (SVs) for chip and analyzer calibration and analytical quantification; 1-3. Non-TIRF fluorescence microscope as a low-cost analyzer and automated software module for fast and user-friendly image acquisition and analysis. Specific Aim 2: Multi-site Technology Validation for Pancreatic Cancer Diagnosis. 2-1. Comparison with qRT-PCR, NGS and ELISA for performance, cost, assay time, and user friendliness. 2-2. Multi-site blind validation: Lab-to-lab repeatability and blind validation comparison at OSU and MSKCC using patient and normal donor serum samples.