ViraLite: An Ultracompact HIV Viral Load Self-Testing System with Internal Quality Control

Abstract

The availability of effective antiretroviral therapy has made HIV manageable, provided patients have consistent access to routine viral load (VL) testing. Nonetheless, access to frequent VL testing remains limited. There is a need for accessible, user-friendly testing systems that allow people living with HIV (PLHIV) to monitor their VL more frequently and empower self-management. Here, we developed ViraLite, a sample-to-answer, compact, accessible, and battery-powered system for HIV viral load monitoring. The system is built upon a probe-based RT-LAMP assay that allows for multiplexed detection and quantification. An internal quality control targeting the RNase P was incorporated to enhance the reliability of the results. A software-reconfigurable real-time sensing system empowered by machine learning and a smartphone-guided protocol was developed in tandem to analyze the multiplexed assay. We analyzed 45 clinically archived samples using ViraLite and benchmarked our results against qRT-PCR, which showed 21 positive and 23 negative samples. Using our process control, ViraLite first identified 17 inconclusive samples that would otherwise be classified as negative. Then, ViraLite classified 14 out of 15 HIV-positive samples (93.3%) and 13 out of 13 HIV-negative samples (100%). The incorporation of RNase P as a process control increased the sensitivity of ViraLite from 66.66% to 93.33%, while maintaining a high specificity (100%). To assess the acceptance of ViraLite among PLHIV, we recruited 480 participants from online and three clinical sites to complete a survey. Over 86% of participants indicated ViraLite had benefits in convenience and privacy, on the other hand 61% of participants indicated concerns with test accuracy. The integration of compact hardware, a reliable assay, and smartphone guidance provides an accurate, easy to use system for PLHIV to self-manage their viral load and update their prescriptions frequently.