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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的移动生物传感