Rapid Sample Preparation and Ultrasensitive Detection of COVID-19 in Human Saliva

人类唾液中 COVID-19 的快速样品制备和超灵敏检测

• 批准号: 10631968
• 负责人: Tak Sing Wong
• 金额: $ 18.89万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10631968
• 关键词: 2019-nCoV; Antibodies; Biological Markers; Biological Models; COVID-19; COVID-19 detection; COVID-19 patient; COVID-19 screening; COVID-19 test; Cessation of life; Coffee; DNA; Detection; Diagnostic tests; Disease Outbreaks; Engineering; Enzyme-Linked Immunosorbent Assay; Goals; Hour; Human; Human Resources; Libraries; Measures; Methods; Molecular Target; Nature; Patients; Performance; Persons; Polymerase Chain Reaction; Preparation; RNA; Rapid screening; Reagent; Research; Reverse Transcription; Saliva; Sampling; Sorting; Specificity; Speed; Surface; Testing; Time; Training; United States; Viral; Viral Antibodies; Virion; Virus; antibody test; antigen test; aptamer; convolutional neural network; cost; deep learning; design; detection limit; detection platform; disorder control; multiplex detection; nanoparticle; neural network algorithm; novel; pandemic disease; particle; post SARS-CoV-2 infection; targeted biomarker; viral RNA; viral detection

Project Summary The coronavirus disease 2019 (COVID-19) – caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) – is a global pandemic with worldwide cases of over 35 million and a death toll of over 1 million people. In the United States, there are a total of over 7.5 million cases with over 210,000 deaths as of October 2020. The ability to identify COVID-19 infected patients rapidly, accurately, and cost-effectively is of paramount importance to control the disease outbreak. The two different types of COVID-19 tests are diagnostic tests and antibody tests. Specifically, the diagnostic tests can only identify active COVID-19 infected patients; the antibody tests can only indicate prior COVID-19 infections but not active cases. These diagnostic tests include quantitative reverse transcription polymerase chain reaction (qRT-PCR) and rapid antigen tests. In particular, the qRT-PCR is currently the most sensitive method (down to ~100 copies/mL) but can take >12 hours to perform and require trained personnel, specific reagents, and expensive machines. The rapid antigen tests provide results within 30 minutes, but these tests require the samples to contain high virus concentration (i.e., >107 virus particles/mL) leading to increasing chances of false-negative cases. In addition, one common example of the antibody tests is the enzyme-linked immunosorbent assay (ELISA). This test only applies to patients who have already developed the antibodies, which could take several days to weeks. Thus far, no single tests can detect both active and previously COVID-19 infected cases. The objective of the proposed research is to develop a proof-of-concept rapid and ultrasensitive virus-detection platform termed viral-ID, which is capable of simultaneous detecting the SARS-CoV-2 RNA and antibodies inside saliva for the identification of both active and previous COVID-19 infections within a single test. Specifically, the viral-ID platform will allow for specific molecular targeting, separation, enrichment, and filtering, all carried out within a single saliva droplet utilizing the nature-inspired coffee ring effect. Targeting and detection of the viral biomarkers will be carried out by aptamer-functionalized nanoparticles and deep-learning-enabled surface- enhanced Raman scattering (SERS), respectively. Using synthetic SARS-CoV-2 RNA and antibody-spiked saliva as a model system, the goal of this research is to demonstrate a bio-analysis platform capable of rapid sample preparation and ultrasensitive detection of both viral RNA (~10 – 100 copies/mL) and antibodies (~femtomolar) in a saliva droplet within 30 minutes. If successful, our viral-ID platform will provide all-in-one and best-in-class diagnostic and antibody test in terms of speed, sensitivity, and specificity for the rapid screening of COVID-19.

项目摘要 2019冠状病毒病（COVID-19）-由新型严重急性呼吸系统综合征引起 冠状病毒2（SARS-CoV-2）-是一种全球性大流行病，全球病例超过3500万，死亡人数为 超过一百万人。在美国，共有超过750万例病例，超过21万人死亡 截至2020年10月。 快速、准确和经济高效地识别COVID-19感染患者的能力至关重要 控制疾病爆发的重要性。两种不同类型的COVID-19检测是诊断检测， 抗体测试具体而言，诊断测试只能识别活动性COVID-19感染患者;抗体 测试只能表明先前的COVID-19感染，而不是活动病例。这些诊断测试包括定量 逆转录聚合酶链反应（qRT-PCR）和快速抗原检测。特别是，qRT-PCR 是目前最灵敏的方法（低至~100拷贝/mL），但可能需要>12小时才能执行， 训练有素的人员、特定的试剂和昂贵的机器。快速抗原检测在30分钟内提供结果。 分钟，但是这些测试要求样品含有高病毒浓度（即，>107个病毒颗粒/mL） 导致假阴性病例的机会增加。此外，抗体测试的一个常见例子 是酶联免疫吸附试验（ELISA）。这项测试只适用于已经 产生抗体可能需要几天到几周的时间。到目前为止，还没有一个单一的测试可以检测到这两个 活跃和先前感染COVID-19的病例。 这项研究的目的是开发一种概念验证的快速和超灵敏的病毒检测方法 一个名为viral-ID的平台，能够同时检测SARS-CoV-2 RNA和内部抗体 唾液用于在一次测试中识别活动性和既往COVID-19感染。具体而言是 病毒识别平台将允许进行特异性分子靶向、分离、富集和过滤， 在一个单一的唾液液滴利用自然灵感的咖啡环效果。病毒的靶向和检测 生物标志物将由适体功能化的纳米颗粒和深度学习功能的表面进行， 增强拉曼散射（Sers）。使用合成的SARS-CoV-2 RNA和抗体 唾液作为一个模型系统，这项研究的目标是证明一个生物分析平台，能够快速 病毒RNA（~10 - 100拷贝/mL）和抗体的样品制备和超灵敏检测 （~飞摩尔）。如果成功，我们的病毒识别平台将提供一体化的， 在快速筛查的速度、灵敏度和特异性方面， 2019冠状病毒病。