Collaborative Research: Ultrasensitive Cancer Biomarker Detection on Biophotonic Chips

1159453/1159423 Schmidt/Hawkins该提案的主要目标是开发一种芯片，该芯片将微流体与依赖于液芯波导的灵敏荧光激发和检测平台相结合。这种芯片实验室设计用于高度灵敏和特异性地检测癌症患者体液中浓度升高的无细胞核酸（CNA）。预期所提出的技术的高灵敏度将消除当前进行CNA的传统PCR测定时所需的扩增的需要，这反过来增加了与该方法相关的复杂性和成本。研究人员建议首先开发一种用于荧光激发的多模干涉方法，以提高信噪比，并实现光谱复用，以提高灵敏度和特异性。第二步涉及将硅光子层与标准PDMS微流体层集成，用于样品制备和过滤。最初将使用针对与高危人乳头瘤病毒（HPV）18相关的病毒核酸的荧光信标对集成设备进行测试。在最后一步中，将使用来自5名黑色素瘤患者、一名健康人和一名白血病患者的血液对该平台进行测试。聚合酶链反应（PCR）方法是目前无细胞核酸（CNA）检测的金标准，它依赖于扩增基因组材料以产生足够大的信号用于光学读出。PI建议展示和验证用于无细胞DNA的多重、无扩增分析的生物光子平台。其核心将是液芯光波导，其在临床相关浓度下对DNA/RNA的单个荧光分子具有检测极限。该项目将解决检测系统各个层面的关键创新，如果成功，将为癌症诊断创造变革性的一步，从而实现对无细胞DNA生物标志物组的非侵入性和定量分析。

1159453/1159423Schmidt/HawkinsThe main goal of the proposal is to develop a chip that combines microfluidics with a sensitive fluorescence excitation and detection platform that relies on liquid-core waveguides. This lab-on-a-chip is designed for highly sensitive and specific detection of cell-free nucleic acids (CNAs) that are found in elevated concentrations within the bodily fluids of cancer patients. The high sensitivity of the proposed technology is expected to obviate the need for amplification that is currently required when performing traditional PCR assays for CNAs, which in turn increase the complexity and cost associated with this approach. The investigators propose to initially develop a multi-mode interferometric approach for fluorescence excitation in order to enhance the signal to noise ratio and to enable spectral multiplexing in order to enhance sensitivity and specificity. The second step involves integration of the silicon photonic layer with standard PDMS microfluidics layers for sample preparation and filtering. The integrated device will be tested initially using fluorescent beacons against viral nucleic acids associated with high-risk human papilloma virus (HPV) 18. In the final step, the platform will be tested using blood from 5 melanoma patients, a healthy human and a leukemia patient.Polymerase chain reaction (PCR) methods are the current gold standard in cell-free nucleic acids (CNA) detection and rely on amplifying the genomic material to produce a sufficiently large signal for optical readout.. The PIs propose to demonstrate and validate a biophotonic platform for multiplexed, amplification-free analysis of cell-free DNA. At its core will be liquid-core optical waveguides that have a limit of detection of single fluorescing molecules of DNA/RNA at clinically relevant concentrations. The project will address key innovations at all levels of the detection system, and, if successful, create a transformative step for cancer diagnosis, enabling non-invasive and quantitative analysis of biomarker panels from cell-free DNA.