(307d) A Rapid CRISPR-Based Self-Testing Platform for Early Detection of HIV

According to the World Health Organization (WHO), an estimated 39.9 million people were living with Human Immunodeficiency Virus (HIV) at the end of 2023, including 1.2 million in the United States. Acute HIV infection, the earliest stage of HIV, typically develops within 2-4 weeks post-exposure, during which the virus rapidly multiplies and spreads. Therefore, early detection is critical to improving individual health and lowering transmission rates. The Centers for Disease Control and Prevention states that HIV can be detected as early as 10 days of post-infection using nucleic acids tests, compared to 18 days for antibody-based tests. Although there are several commercially available nucleic acid tests for HIV approved by the Federal Drug Administration (FDA), they are high-complexity lab-based tests requiring expensive reagents and equipment increasing cost and limiting accessibility for widespread screening. The only FDA-approved in-home HIV test, OraQuick, detects HIV through an antibody-based test. However, there are currently no FDA-approved nucleic acid-based point-of-care or self-testing kits for HIV detection.

This project aims to develop a non-invasive, rapid, simple, and specific self-testing kit using clustered regularly interspaced short palindromic repeats (CRISPR) technology. CRISPR-based diagnostics, coupled with microfluidic platforms, offer great potential for point-of-care testing. Our approach involves extracting HIV-1 RNA from endogenous samples such as whole blood (WB), serum, or plasma paired with RT-LAMP (reverse transcription loop-mediated isothermal amplification) and CRISPR-based detection, which generates a fluorescence signal for diagnosis within a microfluidic device. Using our lysis techniques, we can detect glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an endogenous control RNA from healthy WB at room temperature. Using GAPDH to measure efficiency of our lysis and RNA extraction methods, we are applying our techniques to extract HIV-1 RNA from HIV-WB patient samples at room temperature and further engineer CRISPR to improve our overall sensitivity and specificity of detection. We ultimately seek to deliver a robust strategy to enhance the efficiency and accuracy of our rapid HIV diagnosis platform to serve as a global HIV detection solution.