Abstract
Effective monitoring of HIV treatment remains constrained by limited access to affordable, sensitive, and user-friendly viral load diagnostics, particularly in resource-limited settings. To address this critical gap, a fully automated, point-of-care device was developed that integrates microfluidic sample processing with ultrasensitive bioluminescence-based viral load detection. Upon simple sample loading, the system automates all steps, capturing multiple HIV subtypes with antibody-coated magnetic beads without fold-change, amplifying bioluminescence via enzyme cascades, and quantifying viral load using a low-cost optical sensor. Analytical validation using 87 HIV-spiked plasma samples confirmed a 95 copies per mL detection limit and reliable quantification of viral loads. Clinical testing with 53 patient samples demonstrated 95% sensitivity and 100% specificity, exceeding the performance of currently available POC assays. The assay can be completed within 65 minutes at a cost of less than $3 per test, further supporting its feasibility for widespread implementation. The platform's ease of use and robust performance across different user experience levels support its potential for expanding access to ART monitoring and improving HIV management worldwide.