Towards Rapid Measurement of Non-Nucleoside Reverse Transcriptase Inhibitors in a Portable, Low-Cost Platform

Abstract

While antiretrovirals (ARV) for HIV prevention and treatment have become very effective over the last 40 years of development, many people living with HIV (PLWH) encounter inadequately suppressed viral loads and treatment failure. Providing drug-level feedback (DLF) can identify those with subtherapeutic ARV levels quickly to promote interventions to improve outcomes. Most DLF approaches measure drug levels via liquid chromatography-tandem mass spectrometry (LC-MS/MS), which can incur high costs and lengthy delays that make it unsuitable for near point-of-care settings. In response, we developed a rapid enzymatic assay for the measurement of non-nucleoside reverse transcriptase inhibitors (NNRTIs) based on inhibition of DNA synthesis using an inexpensive, portable instrument. This assay is entitled the REverse transcriptase ACTivity (REACT) assay. This work demonstrates proof-of-concept measurement of multiple NNRTIs in buffer and spiked plasma samples. We also demonstrate the feasibility of using a portable reader to support routine drug measurement at or near the point of care, providing prompt feedback to facilitate rapid interventions.