Rapid, quantitative isothermal molecular assay for POC HIV-1 viral load monitoring using amplification nucleation site analysis

使用扩增成核位点分析进行 POC HIV-1 病毒载量监测的快速定量等温分子测定

• 批准号: 10759148
• 负责人: Jonathan D. Posner
• 金额: $ 23.33万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10759148
• 关键词: Address; Adherence; Biological Assay; Buffers; Characteristics; Chemistry; Clinic; Clinical Treatment; Complex; DNA; Data; Decentralization; Diagnostic; Emulsions; Event; Failure; Genomics; HIV; HIV Infections; HIV drug resistance; HIV-1; Health; Heating; Heterogeneity; Home; Image Analysis; Incubated; Individual; Kinetics; Laboratories; Machine Learning; Measures; Membrane; Methods; Microfluidic Microchips; Molecular; Monitor; Nucleic Acid Amplification Tests; Nucleic Acids; Outcome; Paper; Patients; Persons; Plasma; Poisson Distribution; Polymerase; Porosity; Procedures; Property; Public Health; RNA; Reaction; Resources; Risk; Running; Scheme; Site; Temperature; Testing; Time; Viral; Viral Load result; Viral load measurement; Viscosity; antiretroviral therapy; base; clinically relevant; cost; detection limit; diagnostic platform; diagnostic tool; digital; improved; instrument; isothermal amplification; novel; point-of-care diagnostics; recombinase; response; self testing; statistics; transmission process; viral RNA

Project Summary Over 20 million people living with HIV (PLHIV) are receiving antiretroviral therapy and require HIV viral load testing to identify cases of virological failure and provide actionable information to guide alternative clinical treatment. Current methods for HIV viral load measurement rely on quantitative PCR (qPCR) or digital PCR (dPCR), which are commonly restricted to highly resourced central laboratories and there is a need to decentralize HIV viral load monitoring to enable rapid, clinic-based or home self-testing viral load measurements. We have identified a novel method for implementing digital isothermal amplification that leverages the characteristic viscous reaction buffer of recombinase polymerase amplification (RPA) isothermal amplification chemistry and commercially available porous membranes. We propose to apply amplification nucleation site analysis (ANSA) for HIV-1 viral load monitoring to accurately quantify HIV-1 RNA over clinically relevant HIV-1 subtypes and viral loads. We propose two exploratory aims to demonstrate that ANSA can achieve the required viral load dynamic range and quantitative precision across HIV-1 subtypes.

项目概要 超过 2000 万艾滋病毒感染者 (PLHIV) 正在接受抗逆转录病毒治疗并需要检测艾滋病病毒载量 进行测试以确定病毒学失败的病例并提供可操作的信息来指导替代临床 治疗。目前的 HIV 病毒载量测量方法依赖于定量 PCR (qPCR) 或数字 PCR （dPCR），通常仅限于资源丰富的中心实验室，因此需要 分散 HIV 病毒载量监测，以实现快速、基于诊所或家庭的自测病毒载量测量。 我们已经确定了一种实现数字等温扩增的新方法，该方法利用 重组酶聚合酶扩增 (RPA) 等温扩增的特征粘性反应缓冲液 化学和市售多孔膜。我们建议应用扩增成核位点 用于 HIV-1 病毒载量监测的分析 (ANSA)，以准确量化临床相关 HIV-1 上的 HIV-1 RNA 亚型和病毒载量。我们提出两个探索性目标来证明 ANSA 能够实现所需的目标 HIV-1 亚型的病毒载量动态范围和定量精度。