A novel REverSe Transcriptase Chain Termination (RESTRICT) assay for near-patient, objective monitoring of long-term PrEP adherence

摘要 由富马酸替诺福韦和恩曲他滨组成的口服暴露前预防(PrEP)是有效的 用于预防艾滋病毒感染，但PrEP的疗效高度依赖于药物依从性。在几次试验中 实施研究表明，PrEP客户端难以保持足够的遵从性和持久性， 监测他们的PrEP使用情况是具有挑战性的。预科提供者依赖于自我报告的遵从性，这通常是 不准确和不可靠的。缺乏客观的PrEP遵守情况监测工具导致效率低下 咨询和糟糕的支持性护理。为了解决这些问题，我们完成了一项随机药物动力学研究 在受控直接观察PrEP中确定药物浓度的研究。长期代谢物，如 替诺福韦-二磷酸(TFV-DP)(~17天半衰期)，提供了更准确的长期PrEP图像 坚持不懈。我们最近开发了一种新的酶分析方法，可以半定量地测量浓度 通过检测逆转录酶的抑制来检测TFV-DP的作用，逆转录酶是口服PrEP药物的细胞靶点。在这 提案，我们的主要目标是优化逆转录链终止(限制) 用于测量PrEP客户的药物浓度的化验，以在实施时建立CLIA标准的有效性 在一个近患者的临床实验室中，并评价使用限制性试验的可行性和可接受性 PrEP客户和供应商之间的药物水平测量。我们将用三个例子来检验我们的中心假设 具体目标：(1)校正和优化TFV-DP长效药物限制法 浓度，与金标准液相色谱串联质谱(LC-MS/MS)比较 测量；(2)验证限制性试验是否符合已建立的CLIA标准，以使临床能够 报告用于监测TFV-DP长期药物浓度的客观近患者测量方法；(3) 在PrEP服务对象和提供者中评估近患者TFV-DP检测的可行性和可接受性 西雅图主要的PrEP诊所。我们提出的研究将是第一个验证快速、近距离长期治疗的研究 口服PrEP依从性的客观测量。这项研究还将提供有关可行性和 改进PrEP交付和监测以预防艾滋病毒传播的新方法的可接受性。这个 这项研究的结果将开发一种新的工具，可能有助于改善美国和世界各地的PrEP交付。

ABSTRACT Oral pre-exposure prophylaxis (PrEP), composed of tenofovir disoproxil fumarate and emtricitabine, is effective for preventing HIV acquisition, but PrEP efficacy is highly dependent on drug adherence. In several trials and implementation studies, PrEP clients have difficulties maintaining adequate adherence and persistence, and monitoring their PrEP use is challenging. PrEP providers have relied on self-reported adherence, which is often inaccurate and unreliable. The lack of an objective PrEP adherence monitoring tool has led to inefficient counseling and poor supportive care. To address these issues, we completed a randomized pharmacokinetic study to determine drug levels during controlled directly-observed PrEP. Longer-term metabolites, such as tenofovir-diphosphate (TFV-DP) (~17-day half-life), provide a more accurate picture of long-term PrEP adherence. We recently developed a novel enzymatic assay that semi-quantitatively measures the concentration of TFV-DP by measuring inhibition of reverse transcriptase, which is the cellular target of oral PrEP drugs. In this proposal, our primary objectives are optimizing the REverSe TRanscrIptase Chain Termination (RESTRICT) assay to measure drug concentrations of PrEP clients, to establish validation for CLIA criteria when implemented in a near-patient clinical lab, and to evaluate the feasibility and acceptability of using the RESTRICT assay for drug level measurement among PrEP clients and providers. We will test our central hypotheses with three specific aims: (1) to calibrate and optimize the RESTRICT assay for measuring long-term TFV-DP drug concentrations, compared to gold-standard liquid chromatography tandem mass spectrometry (LC-MS/MS) measurement; (2) to validate the RESTRICT assay for meeting established CLIA criteria to enable clinical reporting of an objective near-patient measure for monitoring long-term TFV-DP drug concentrations; (3) to evaluate the feasibility and acceptability of near-patient TFV-DP testing among PrEP clients and providers at a major PrEP clinic in Seattle. Our proposed study will be the first to validate a rapid, near-patient long-term objective measure of oral PrEP adherence. This study will also provide crucial data on the feasibility and acceptability of a novel approach for improving PrEP delivery and monitoring to prevent HIV transmission. The results of this study will develop a new tool that may help improve PrEP delivery in the US and worldwide.