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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

基本信息

批准号：
10458559
负责人：
Changchun Liu
金额：
$ 45.98万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
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：
2020-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10458559
关键词：
AIDS/HIV problem 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 Data Reporting Detection Developing Countries Development Devices Early Diagnosis Early Intervention Engineering Ensure Environment Epidemic Equipment Evaluation Fingers Fluorescence HIV HIV Infections HIV diagnosis HIV-1 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 Research Resource-limited setting Reverse Transcriptase Polymerase Chain Reaction Reverse Transcription Risk Role Sampling Sensitivity and Specificity Signal Transduction Statistical Data Interpretation System Technology Test Result Testing Time Training Treatment Effectiveness Treatment Failure Universities Validation Viral Viral Load result Viremia Whole Blood Zambia antiretroviral therapy base 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

项目摘要

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.

自供电样品浓缩和基于CRISPR的移动的生物传感 HIV-1 RNA检测摘要艾滋病毒/艾滋病已成为一个主要的公共卫生问题，影响到全世界约3790万人。早期诊断在血清转换窗口期的急性HIV感染率将有助于早期干预。抗逆转录病毒治疗期间艾滋病毒感染者的抗逆转录病毒治疗（ART），需要经常监测艾滋病毒载量，以确认治疗有效性，并确定病毒反弹。HIV病毒载量检测，定量HIV病毒RNA（循环HIV 在血浆中检测HIV（病毒）是ART监测和急性HIV检测的最准确和可靠的方法。然而，目前标准的HIV病毒载量检测方法依赖于昂贵的设备和训练有素的人员，限制了其在集中实验室和医院环境中的临床应用。市售的基于免疫测定的床旁（POC）诊断技术，如OraQuick 艾滋病毒自我检测（HIVST）不能有效检测急性艾滋病毒感染以及ART失败。作为因此，缺乏一种简单、快速、负担得起的用于HIV RNA检测的POC诊断工具，这些人不了解自己的病情，妨碍及时接受抗逆转录病毒治疗。为了填补这一空白，我们建议开发一种低成本（~ 5美元）、快速（< 35分钟）、敏感（<1,000拷贝/mL）、定期聚类的间隔短回文重复序列（CRISPR）生物传感平台，用于使用手指检测HIV病毒载量全血采血量（约50 µL）。在R61阶段（目标1-3），我们将：i）高度开发和优化用于下一代基于核酸的分子的灵敏且特异的CRISPR生物传感技术 ii）设计和制造一次性“血液应答”CRISPR生物传感装置，集成了自供电血浆分离、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的集成CRISPR生物传感的性能平台，并通过在美国测试HIV临床样本，严格验证其临床应用的可行性和赞比亚如果成功，这种简单，快速，负担得起的POC检测平台将使急性艾滋病毒在家里进行诊断和病毒载量检测，并适用于艾滋病毒感染最多的资源有限的环境普遍存在。