Self-Powered Sample Concentrating and CRISPR-based Biosensing for Moile HIV-1 RNA Detection

用于 Moile HIV-1 RNA 检测的自供电样本浓缩和基于 CRISPR 的生物传感

• 批准号: 10878025
• 负责人: Changchun Liu
• 金额: $ 83.01万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10878025
• 关键词: Acute; Adopted; Affect; Bioinformatics; Biological Assay; Biosensing Techniques; Blinded; Blood; Blood capillaries; Blood specimen; Cellular Phone; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Country; Cytolysis; Data; Detection; Developing Countries; Development; Devices; Early Diagnosis; Early Intervention; Engineering; Ensure; Environment; Epidemic; Equipment; Evaluation; Fingers; Fluorescence; HIV; HIV Infections; HIV diagnosis; HIV-1; HIV/AIDS; Health; Home; Hospitals; Human Resources; Human immunodeficiency virus test; Immunoassay; Individual; Infant; Infectious Diseases Research; International; Laboratories; Membrane; Methods; Microfluidic Microchips; Monitor; Mothers; Nucleic Acid Amplification Tests; Nucleic Acids; Patients; Pennsylvania; Performance; Persons; Phase; Plasma; Play; Polymerase; Public Health; RNA; Reaction; Recovery; Reporting; Research; Resource-limited setting; Reverse Transcriptase Polymerase Chain Reaction; Reverse Transcription; Risk; Role; Sampling; Signal Transduction; Specificity; Statistical Data Interpretation; System; Technology; Test Result; Testing; Training; Treatment Effectiveness; Treatment Failure; Universities; Validation; Viral; Viral Load result; Viremia; Virus; Whole Blood; Zambia; antiretroviral therapy; biomaterial compatibility; clinical application; cost; design; detection platform; detection sensitivity; diagnostic technologies; diagnostic tool; field study; improved; innovation; instrument; instrumentation; internal control; isothermal amplification; molecular diagnostics; multidisciplinary; next generation; point of care; point-of-care detection; point-of-care diagnostics; portability; recombinase; self testing; seroconversion; tool; transmission process; viral RNA; viral detection; viral rebound

Self-Powered Sample Concentrating and CRISPR-based Biosensing for Mobile HIV-1 RNA Detection Abstract HIV/AIDS has become a major public health concern affecting ~37.9 million people worldwide. Early diagnosis of acute HIV infection during seroconversion window will facilitate early intervention. During antiretroviral treatment (ART) of HIV-infected patients, it requires frequent monitoring of HIV viral load to confirm treatment effectiveness, and to identify viral rebound. HIV viral load testing that quantifies HIV viral RNA (circulating HIV virus) in plasma is the most accurate and reliable approach for the ART monitoring and acute HIV detection. However, current standard HIV viral load testing methods rely on expensive equipment and well-trained personnel, limiting their clinical applications in centralized laboratories and hospital environments. Commercially available immunoassay-based point of care (POC) diagnostic technologies, such as OraQuick® HIV Self-Test (HIVST), are not effective to detect acute HIV infections, as well as ART failure. As a consequence, the lack of a simple, rapid, affordable, POC diagnostic tool for HIV RNA detection leaves many individuals unaware of their condition and impedes timely antiretroviral treatment. To fill this gap, we propose to develop a low-cost (~ $ 5), rapid (< 35 min), and sensitive (<1,000 copies/mL), clustered regularly interspaced short palindromic repeats (CRISPR) biosensing platform for HIV viral load testing using finger- prick volume (~50 µL) of whole blood. In the R61 phase (Aims 1-3), we will: i) develop and optimize highly sensitive and specific CRISPR biosensing technology for next-generation nucleic acid-based molecular diagnostics, and ii) design and fabricate a disposable "blood-to-answer", CRISPR biosensing device that integrates self-powered plasma separation, viral RNA enrichment, and CRISPR biosensing detection. In the Phase 33 (Aims 4-5), we will systematically evaluate the performance of our integrated CRISPR biosensing platform, and rigorously validate its feasibility for clinical application by testing HIV clinical samples in the US and Zambia. If successful, such a simple, rapid, affordable, POC detection platform will enable acute HIV diagnosis and viral load testing at home and be appropriate for resource-limited settings where HIV is most prevalent.

自动样本集中和基于CRISPR的移动生物传感 HIV-1 RNA检测 抽象的 艾滋病毒/艾滋病已成为影响全世界约3790万人的主要公共卫生问题。早期诊断 血清转化窗口期间急性HIV感染将有助于早期干预。在抗逆转录病毒期间 HIV感染患者的治疗（ART），需要经常监测HIV病毒负荷以确认治疗 有效性，并确定病毒反弹。 HIV病毒载荷测试该量HIV病毒RNA（循环HIV 血浆中的病毒）是ART监测和急性HIV检测的最准确，最可靠的方法。 但是，当前的标准HIV病毒负荷测试方法依赖于昂贵的设备和训练有素 人员，限制其在集中实验室和医院环境中的临床应用。 基于市售的免疫测定点护理点（POC）诊断技术，例如Oraquick® 艾滋病毒自我测试（HIVST）无效检测急性艾滋病毒感染以及艺术失败。作为 结果，缺乏简单，快速，负担得起的POC诊断工具用于HIV RNA检测有许多 个人不知道自己的状况并阻碍及时的抗逆转录病毒治疗。为了填补这一空白，我们提出了 要开发低成本（〜$ 5），快速（<35分钟）和敏感（<1,000份/毫升），请定期聚类 间隔短的短静脉重复序列（CRISPR）生物传感平台，用于使用Finger-finger- 全血的刺量（〜50 µL）。在R61阶段（目标1-3），我们将：i）高度开发和优化 下一代核酸分子的敏感和特定的CRIS PR生物传感技术 诊断和ii）设计和制造一次性的“鲜血降血”，CRISPR生物传感设备 整合了自动血浆分离，病毒RNA富集和CRISPR生物传感检测。在 第33阶段（目标4-5），我们将系统地评估我们集成的CRISPR生物传感的性能 平台，并严格验证其对临床应用的可行性，通过测试美国的HIV临床样本 和赞比亚。如果成功的话，如此简单，快速，负担得起的POC检测平台将引起急性HIV 诊断和病毒负载测试在家中，适合于艾滋病毒最多的资源有限设置 流行。