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High Throughput Digital Droplet ELISA for Ultrasensitive Multiplexed Diagnostics

用于超灵敏多重诊断的高通量数字液滴 ELISA

基本信息

批准号：
10359798
负责人：
David Aaron Issadore
金额：
$ 17.42万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-04 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10359798
关键词：
Address Antibodies Attention Benchmarking Benign Biological Assay Biological Markers Cancer Center Cancer Diagnostics Cancer Patient Cellular Phone Clinical Clinical Sensitivity Cloud Computing Code Collaborations Color Complex DNA Detection Development Devices Diagnosis Disease Disease Progression Drug Targeting Electronics Emulsions Enzyme-Linked Immunosorbent Assay Fluorescence Gold Grant Heterogeneity Human Hybrids Individual Industry Laboratories Lasers Lesion Light Liquid substance Malignant - descriptor Malignant Neoplasms Malignant neoplasm of pancreas Measurement Measures Methods Microfluidic Microchips Microspheres Modality Molecular Monitor Nature Neoplasm Metastasis Nucleic Acids Patients Physiological Plasma Population Prognosis Proteins Publishing RNA Reaction Reproducibility Research Resolution Running Sampling Sensitivity and Specificity Source Technology Telecommunications base biomarker panel cancer biomarkers cancer diagnosis chemical reaction circulating biomarkers clinical biomarkers clinical diagnostics cohort cost cytokine detection limit detector digital disease diagnostic disorder control drug efficacy experimental study falls follow-up handheld mobile device human subject improved innovation instrumentation microelectronics microfluidic technology molecular marker multiplex detection multiplex diagnostics personalized medicine point of care portability protein biomarkers single molecule small molecule tool tumor

项目摘要

Abstract: Digital assays — in which ultra-sensitive molecular measurements are made by performing millions of parallel experiments in picoliter droplets — have generated much recent enthusiasm due to their single molecule resolution of RNA, DNA, and proteins, and their robustness to reaction conditions. These assays have enormous untapped potential for point of care disease diagnostics, but are currently mainly confined to laboratory settings due to the cumbersome instrumentation necessary to generate, control, and measure tens of millions of independent droplets. To overcome this challenge, we propose a hybrid microelectronic / microfluidic chip to `unlock' droplet-based assays for clinical use. Our microdroplet megascale detector (µMD) can generate and detect the fluorescence of millions of droplets per second (1000× faster than existing digital approaches), while achieving a 1000x greater sensitivity than conventional ELISA, using only a conventional cell phone camera. The key innovation of our approach is borrowed from the telecommunications industry, wherein we modulate the excitation light with a pseudorandom sequence that enables individual droplets to be resolved that would otherwise overlap due to the limited frame rate of digital cameras. To demonstrate the power of our approach, we focus our attention on the diagnosis of pancreatic cancer.

摘要：数字分析-通过执行超灵敏分子测量，在皮升液滴中进行的数百万次平行实验，由于它们对RNA、DNA和蛋白质的单分子分辨率，以及它们对反应条件这些检测方法在床旁疾病方面具有巨大的未开发潜力诊断，但目前主要局限于实验室设置，由于繁琐的产生、控制和测量数千万个独立的水滴。为了克服这一挑战，我们提出了一种混合微电子/微流体芯片， “解锁”基于液滴的检测以供临床使用。我们的微滴百万级探测器（µMD）可以每秒产生并检测数百万个液滴的荧光（比现有的数字方法），同时实现比传统方法高1000倍的灵敏度 ELISA，只使用传统的手机摄像头。我们方法的关键创新在于从电信行业借来的，其中我们用伪随机序列，其使得能够分辨单独的液滴，否则由于数码相机有限的帧速率，为了证明我们的力量胰腺癌的早期诊断方法有哪些？