ViraLite: an ultracompact HIV viral load self-testing system with internal quality control

Effective antiretroviral therapy has transformed HIV into a manageable chronic condition, provided viral suppression is maintained through routine viral load (VL) monitoring. Access to frequent VL testing remains limited, particularly outside centralized clinical settings. Decentralized and at-home HIV VL testing requires low-volume systems designed for patient operation. These systems must distinguish true viral suppression from test failure. Many approaches lack internal process verification, which makes negative results ambiguous. Here, we present ViraLite, an ultracompact, battery-powered HIV VL monitoring system that integrates reverse transcription loop-mediated isothermal amplification (RT-LAMP) with an RNase P internal process control, machine learning–assisted fluorescence analysis for one-pot multiplexing, and smartphone-guided operation. We evaluated ViraLite using 45 clinically archived plasma samples and benchmarked it against reverse transcription quantitative polymerase chain reaction (RT-qPCR). The internal process control identified 17 inconclusive tests that would otherwise be misclassified as negative. Among valid tests, ViraLite achieved 93.3% sensitivity and 100% specificity versus RT-qPCR. ViraLite enables decentralized HIV VL testing with low sample volume and interpretable negative results. This capability can expand monitoring beyond traditional clinic workflows and addresses a key barrier to patient-operated VL testing.