Pilot Evaluation of an Enzymatic Assay for Rapid Measurement of Antiretroviral Drug Concentrations
| Autor: | Tim R. Cressey, Ashley Bardon, Benjamin Sullivan, Tiffany J. Lo, Jonathan D. Posner, Ayokunle Olanrewaju, Paul K. Drain |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Adult genetic structures Anti-HIV Agents Metabolite 030106 microbiology Short Report Antiretroviral drug HIV Infections Pilot Projects Infectious and parasitic diseases RC109-216 Pharmacology Biology behavioral disciplines and activities 03 medical and health sciences chemistry.chemical_compound Pre-exposure prophylaxis 0302 clinical medicine Liquid chromatography–mass spectrometry Tandem Mass Spectrometry Virology Humans 030212 general & internal medicine Whole blood Enzyme Assays chemistry.chemical_classification Enzymatic assay Reverse transcriptase HIV Reverse Transcriptase Dried blood spot Infectious Diseases Enzyme chemistry nervous system Adherence Drug Monitoring Tenofovir diphosphate psychological phenomena and processes Chromatography Liquid |
| Zdroj: | Virology Journal Virology Journal, Vol 18, Iss 1, Pp 1-6 (2021) |
| Popis: | ObjectiveMaintaining adequate drug adherence is crucial to ensure the HIV prevention benefits of pre-exposure prophylaxis (PrEP). We developed an enzymatic assay for rapidly measuring tenofovir-diphosphate (TFV-DP) concentrations—a metabolite that indicates long-term PrEP adherence.SettingThe study was conducted at the Madison HIV Clinic at Harborview Medical Center in Seattle.MethodsWe enrolled adults receiving standard oral PrEP, and individuals not receiving any antiretrovirals. We measured TFV-DP concentrations in diluted whole blood using our novel REverSe TRanscrIptase Chain Termination (RESTRICT) assay, based on inhibition of HIV reverse transcriptase (RT) enzyme. Blood samples were diluted in water, DNA templates, nucleotides, RT, and intercalating dye added, and results measured with a fluorescence reader—stronger fluorescence indicated higher RT activity. We compared RESTRICT assay results to TFV-DP concentrations from matched dried blood spot samples measured by liquid chromatography tandem mass spectrometry (LC–MS/MS) using ≥ 700 fmol/punch TFV-DP as a threshold for adequate adherence (≥ 4 doses/week).ResultsAmong 18 adults enrolled, 4 of 7 participants receiving PrEP had TFV-DP levels ≥ 700 fmol/punch by LC–MS/MS. RESTRICT fluorescence correlated with LC–MS/MS measurements (r = − 0.845,p ConclusionsThis novel enzymatic assay for measuring HIV reverse transcriptase activity may be suitable for distinguishing TFV-DP concentrations in blood that correspond to protective PrEP adherence. |
| Databáze: | OpenAIRE |
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