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

摘要 由富马酸替诺福韦酯和恩曲他滨组成的口服暴露前预防（PrEP）有效 预防HIV感染，但PrEP的疗效高度依赖于药物依从性。在几次审判中， 实施研究，PrEP客户难以保持足够的依从性和持久性， 监测他们的PrEP使用是一项挑战。PrEP提供者依赖于自我报告的依从性，这通常是 不准确和不可靠。缺乏客观的PrEP依从性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代动力学研究。 研究，以确定药物水平在控制直接观察的PrEP。长期代谢物，如 替诺福韦二磷酸盐（TFV-DP）（~17天半衰期），提供更准确的长期PrEP图片 坚持。我们最近开发了一种新的酶测定，半定量测量浓度 通过测量逆转录酶的抑制作用来测定TFV-DP，逆转录酶是口服PrEP药物的细胞靶点。在这 我们的主要目标是优化REverSe转肽酶链终止（RESTRICT） 测定PrEP客户的药物浓度，以在实施时验证CLIA标准 在接近患者的临床实验室中，并评价使用RESTRICT检测试剂盒进行 PrEP客户和供应商之间的药物水平测量。我们将用三个假设来检验我们的中心假设。 具体目的：（1）校准和优化用于测量长期TFV-DP药物的RESTRICT试验 浓度，与金标准液相色谱串联质谱法（LC-MS/MS）相比 （2）验证RESTRICT测定是否符合既定的CLIA标准， 报告用于监测长期TFV-DP药物浓度的客观近患者测量值;（3） 评估在PrEP客户和提供者中进行近患者TFV-DP检测的可行性和可接受性， 西雅图的一家主要的预防性健康教育诊所。我们提出的研究将是第一个验证快速，近病人的长期 口服PrEP依从性的客观测量。这项研究还将提供关于可行性的重要数据， （c）改进PrEP提供和监测以预防艾滋病毒传播的新方法的可接受性。的 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提供。

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.