Collaborative Research: Ultrasensitive Cancer Biomarker Detection on Biophotonic Chips

1159453/1159423施密特/霍金斯该提案的主要目标是开发一种将微流体与依赖液芯波导的灵敏荧光激发和检测平台相结合的芯片。这种芯片实验室设计用于对癌症患者体液中浓度升高的无细胞核酸(CNA)进行高灵敏度和特异性的检测。拟议技术的高灵敏度预计将消除目前在对CNA进行传统的PCR分析时所需的扩增需求，这反过来又增加了与这种方法相关的复杂性和成本。研究人员建议初步开发一种用于荧光激发的多模干涉方法，以提高信噪比，并实现光谱多路复用，以提高灵敏度和特异性。第二步是将硅光子层与标准的PDMS微流体层进行集成，以进行样品制备和过滤。集成设备最初将使用与高危人乳头瘤病毒(HPV)18相关的病毒核酸的荧光信标进行测试。在最后一步，该平台将使用来自5名黑色素瘤患者、一名健康人和一名白血病患者的血液进行测试。聚合酶链式反应(PCR)方法是目前无细胞核酸(CNA)检测的金标准，它依赖于放大基因组材料来产生足够大的信号用于光学读出。PI建议演示和验证用于多路、无扩增的无细胞DNA分析的生物光子学平台。它的核心将是液芯光波导，在临床相关浓度下，它对单一荧光分子DNA/RNA的检测能力是有限的。该项目将涉及检测系统各级的关键创新，如果成功，将为癌症诊断创造一个变革性的步骤，实现对来自无细胞DNA的生物标记物面板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.