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：
9889673
负责人：
David Aaron Issadore
金额：
$ 21.33万
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 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9889673
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 Diagnostic 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 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 detector digital disorder control drug efficacy experimental study falls follow-up handheld mobile device human subject improved innovation instrumentation microfluidic technology molecular marker multiplex detection outcome forecast 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.

抽象的：数字测定 - 通过执行超敏感的分子测量在Picoliter液滴中进行了数百万平行的实验 - 产生了许多最近的热情由于它们的RNA，DNA和蛋白质分子分辨率及其鲁棒性反应条件。这些测定具有巨大的未开发的护理疾病潜力诊断，但目前主要仅限于实验室环境生成，控制和测量数千万独立的仪器飞沫。为了克服这一挑战，我们提出了一个混合微电子 /微流体芯片 “解锁”基于临床用途的基于液滴的测定法。我们的微圆头大型巨型探测器（µMD）可以生成和检测每秒数百万滴的荧光（比1000倍现有的数字方法），同时获得比传统的敏感性高1000倍 ELISA，仅使用常规手机摄像头。我们方法的关键创新是从电信行业借来的伪随机序列可以解决单个液滴的解决由于数码相机的帧速率有限，重叠。展示我们的力量方法，我们将注意力集中在诊断癌上。