Plasmon-enhanced Lateral Flow Assay for Multiplexed Detection of SARS-CoV-2 RNA and Antigens in Point-of-Care Settings

等离激元增强侧流分析用于在护理点环境中多重检测 SARS-CoV-2 RNA 和抗原

• 批准号: 2224610
• 负责人: Srikanth Singamaneni
• 金额: $ 48万
• 依托单位: Washington University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-11-01 至 2025-10-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2224610&HistoricalAwards=false
• 关键词: Plasmon; enhanced; Lateral; Flow; Assay

Lateral flow assays (LFAs) are amongst the simplest, fastest, and cheapest point-of-care (POC) and at-home biodiagnostic platforms and offer a broad potential for individualized and population-level screening for infectious diseases. Conventional LFAs rely on gold nanoparticles as labels to produce visually detectable color in the presence of the target biomarker. However, the sensitivity of LFAs relying on gold nanoparticles as labels is often insufficient to detect low concentrations of biomarkers, resulting in false negatives at the early stage of disease progression. This project seeks to explore the use of an ultrabright fluorescent label for improving the sensitivity of the LFA by more than 100-fold. The project also aims to design and build a personal handheld fluorimeter that will communicate wirelessly with a linked mobile device to measure the fluorescence signal from an LFA test strip. Harnessing the ultrabright nanolabel and the personal handheld fluorimeter, a novel diagnostic platform that simultaneously detects SARS-CoV-2 (the virus causing COVID-19) RNA and antigen is envisioned. Simultaneous detection of viral RNA and antigen can potentially improve diagnostic accuracy and accurately indicate the stage of illness. The project also involves the development of a new Nano-Biosensors Summer School activity intended to introduce middle school students to nanotechnology and biosensors, with the goal of stimulating interest in science/engineering while instilling confidence in pursuing these topics among under-represented groups.There is a pressing need for a new generation of biosensor technologies that encompass the combined characteristics of low-cost instrumentation, simple assay methods, high sensitivity, accurate quantitation, and rigorous validity. The ultimate goal of the project is to design and realize a multiplexed plasmon-enhanced fluorescence-based lateral flow assay (p-LFA) for the detection and quantification of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigen and RNA in resource-limited settings. Specific objectives include: (i) Design, realize and validate an ultrasensitive multiplexed p-LFA for simultaneous detection and quantification of SARS-CoV-2 RNA and antigen; (ii) Devise efficient sample processing methods for simultaneous detection of RNA and antigens in nasopharyngeal swabs and saliva from coronavirus disease 2019 (COVID-19) patients; and (iii) Build and test a smart phone-interfaced portable p-LFA reader for deploying the proposed biosensing technology in point-of-care (POC) and resource-limited settings. The project represents a transformative advance in that it seeks to establish a novel biodiagnostic platform with sensitivity exceeding that of the standard laboratory tests, a simple assay workflow compatible with self-testing at the POC and patient home, rapid sample-to-answer, and an inexpensive/portable instrument. If successful, the proposed amplification-free assay enables spatially-multiplexed detection and quantification of SARS-CoV-2 RNA and antigens on the same LFA strip, which shortens the sample-to-answer time and potentially differentiates individuals in the infectious stage of the illness from those who have passed it. Beyond COVID, simultaneous detection of RNA and antigen is broadly applicable for highly specific and sensitive diagnosis of various infectious diseases, including Ebola, Influenza, and Human Immunodeficiency Virus.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

横向流量测定（LFA）是最简单，最快，最便宜的护理点（POC）和在家生物诊断平台，并为传染病的个性化和人口水平筛查提供了广泛的潜力。 传统的LFA依靠金纳米颗粒作为标签，在靶生物标志物存在下产生可检测到的颜色。 但是，依赖金纳米颗粒作为标签的LFA的敏感性通常不足以检测到低浓度的生物标志物，从而在疾病进展的早期阶段导致错误的负面因素。 该项目旨在探索使用Ultrabright荧光标签，以提高LFA的敏感性超过100倍以上。 该项目还旨在设计和构建个人手持式荧光计，该计算机将与链接的移动设备无线通信，以测量LFA测试条的荧光信号。 利用Ultrabright纳米标签和个人手持式荧光计，这是一个新型的诊断平台，同时检测到SARS-COV-2（引起Covid-19的病毒）RNA和抗原。 同时检测病毒RNA和抗原可以潜在地提高诊断准确性，并准确地表明疾病阶段。 The project also involves the development of a new Nano-Biosensors Summer School activity intended to introduce middle school students to nanotechnology and biosensors, with the goal of stimulating interest in science/engineering while instilling confidence in pursuing these topics among under-represented groups.There is a pressing need for a new generation of biosensor technologies that encompass the combined characteristics of low-cost instrumentation, simple assay methods, high灵敏度，准确的定量和严格的有效性。 该项目的最终目标是设计和实现基于荧光荧光的荧光横向流动测定（P-LFA），用于在资源受限的设置中检测和定量严重急性呼吸综合征冠状病毒2（SARS-COV-2）抗原和RNA。 特定目标包括：（i）设计，实现和验证超敏的多重P-LFA，以同时检测和定量SARS-COV-2 RNA和抗原； （ii）设计有效的样品加工方法，用于同时检测2019年鼻咽拭子中的RNA和抗原和唾液中的RNA和唾液； （iii）构建和测试智能手机交织的便携式P-LFA读取器，用于在护理（POC）和资源有限的设置中部署所提出的生物传感技术。 该项目代表了一种变革性的进步，它试图建立一个新型的生物诊断平台，其灵敏度超过了标准实验室测试的敏感性，简单的测定工作流与POC和患者住宅的自我测试兼容，快速样品到招牌，以及便宜的/便携式仪器。 如果成功，则提出的无扩增测定可以使SARS-COV-2 RNA和抗原在同一LFA条上的空间进行检测和量化，从而缩短了样本到移交的时间，并有可能在疾病的传染性阶段中区分个体。 Beyond COVID, simultaneous detection of RNA and antigen is broadly applicable for highly specific and sensitive diagnosis of various infectious diseases, including Ebola, Influenza, and Human Immunodeficiency Virus.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.