Integrated Acoustofluidic Plasmonic Molecular Diagnostic System for Detecting MicroRNA Biomarkers

该项目的目标是开发一种用于快速隔离的集成诊断系统， 人类临床循环异构体微核糖核酸(MiRNA)生物标志物的检测和多重鉴定 用于疾病早期检测的样本。MiRNAs调节失调常见于各种疾病，包括 癌症、心血管疾病和感染。最近的研究还表明，miRNAs可以 通过外体从肿瘤细胞分泌到血流中；它们的表达谱可用于癌症分类 类型。因此，循环的外体miRNAs被认为是有希望的早期检测和治疗的生物标志物。 癌症的诊断。然而，由于外切体分离和miRNA带来的技术困难 对这些小分子的分析、检测尚未应用于临床实践。因此，有必要 开发替代分析策略，可以提供比传统方法更多的优势。在这 项目，我们提出了一个集成的声流控等离子体分子诊断系统，设计用于快速 胞外体miRNA生物标志物的分离和多重检测。跨学科的方法将提供 推进疾病精确临床诊断的新能力。该系统集成了两个独特的 技术：(1)可以高产率和高纯度快速分离外切体的声流控技术；(2) 基于表面增强拉曼散射(SERS)的“反向分子哨兵”(IMS)纳米探针直接检测 MiRNA检测。虽然与结直肠癌相关的miRNAs将被用作模型系统，但 拟议的项目还将导致开发一种普遍适用的护理点诊断技术 用于其他类型的疾病。具体目标是：(1)开发和集成声流控系统 胞外体miRNA的分离；(2)制备多重检测胞外体miRNA的IMS纳米探针 生物标志物；(3)SERS-声流系统的技术评价。我们将在合并后的基础上 我们的跨学科团队拥有建立技术验证所需的知识，以准备 未来在临床环境中使用的技术。在整个项目中，将开发一个综合系统，并 可用于临床样本中多种外体miRNA的原位分析，而不需要miRNA 提取和扩增。在技术验证阶段，将结果与所获得的结果进行比较 使用常规检测方法(如qRT-PCR)。该系统将导致无样品制备和“样品到样品”。 回答“基于现有和测试子组件的集成的分析方法。这是一项新的 诊断系统能够加强早期发现和筛查领域的研究和翻译 除了CRC之外，它还可以检测各种疾病；它非常适合于护理点检测。拟议的系统 代表着在生物医学研究、诊断和筛查方面具有变革潜力的重大创新。

The goal of this project is to develop an integrated diagnostic system designed for rapid isolation, detection, and multiplexed identification of circulating exosomal microRNA (miRNA) biomarkers in human clinical samples for early detection of diseases. Dysregulation of miRNAs is often observed in various diseases including cancer, cardiovascular illnesses, and infection. Recent studies have also demonstrated that miRNAs can be secreted from tumor cells into bloodstream via exosomes; their expression profiles can be used to classify cancer types. Therefore, circulating exosomal miRNAs are considered promising biomarkers for early detection and diagnosis of cancer. However, because of technical difficulties arising from exosome isolation and miRNA analysis, detection of these small molecules has not been adopted into clinical practice. Thus, there is a need to develop alternative analysis strategies that could offer more advantages over conventional methods. In this project, we propose an integrated acoustofluidic plasmonic molecular diagnostic system designed for rapid isolation and multiplexed detection of exosomal miRNA biomarkers. The cross-disciplinary approach will provide new capabilities to advance the precise clinical diagnosis of diseases. This system integrates two unique technologies: (1) acoustofluidic technology that can rapidly isolate exosomes with high yield and purity; and (2) surface-enhanced Raman scattering (SERS)-based “Inverse Molecular Sentinel” (iMS) nanoprobes for direct miRNA detection. Although miRNAs related to colorectal cancer (CRC) will be used as the model system, the proposed project will also lead to the development of a generally applicable point-of-care diagnostic technology for other types of diseases. The specific aims are: (1) Develop and integrate an acoustofluidic system for exosomal miRNA isolation; (2) Develop iMS nanoprobes for multiplexed detection of exosomal miRNA biomarkers; and (3) Technical Evaluation of the SERS-acoustofluidic system. We will build upon the combined knowledge of our interdisciplinary team establishing the technical validation required to ready this integrated technology for future use in clinical settings. Throughout this project, an integrated system will be developed and validated for in-situ analysis of multiple exosomal miRNAs from clinical samples without the need for miRNA extraction and amplification. In the technical validation phase, the results will be compared to those obtained using conventional assays (e.g. qRT-PCR). The system will lead to a no-sample preparation and “sample-to- answer” analysis approach that is based on the integration of existing and tested sub-components. This new diagnostic system is capable of enhancing research and translation in the areas of early detection and screening of various diseases beyond CRC; it is exceedingly well suited for point-of-care testing. The proposed system represents a major innovation with transformative potential in biomedical research, diagnostics, and screening.