Pilot evaluation of an enzymatic assay for rapid measurement of antiretroviral drug concentrations

摘要

Workflow for RESTRICT assay for measuring antiretroviral drug concentrations. Patient blood samples are diluted in water to lyse red blood cells, release intracellular tenofovir diphosphate, and minimize assay interference from blood matrix components. Next diluted blood is added to the RESTRICT assay master mix and incubated at 37 °C for 30 min to allow time for DNA synthesis reactions. Finally, PicoGreen dye is added to stop the reaction and provide fluorescence readout. Samples with low TFV-DP concentration are conducive to DNA synthesis and provide high fluorescence signals, meanwhile samples with high TFV-DP concentration lead to DNA chain synthesis and provide low fluorescence signals