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.

项目摘要 超过2000万艾滋病毒感染者正在接受抗逆转录病毒治疗，并需要艾滋病毒载量 检测，以确定病毒学失败的病例，并提供可操作的信息，以指导替代的临床 治疗目前用于HIV病毒载量测量的方法依赖于定量PCR（qPCR）或数字PCR （dPCR），通常仅限于资源丰富的中心实验室，因此有必要 分散艾滋病毒载量监测，以便能够快速、基于诊所或家庭自测的病毒载量测量。 我们已经确定了一种用于实现数字等温扩增的新方法，该方法利用了 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。我们建议应用放大成核位点 用于HIV-1病毒载量监测的ANSA分析，以准确定量HIV-1 RNA相对于临床相关HIV-1 亚型和病毒载量。我们提出了两个探索性的目标，以证明ANSA可以实现所需的 HIV-1亚型间病毒载量动态范围和定量精确度。