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

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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.