Novel Quantitative Cost-effective Assay for the Diagnosis and Monitoring of HIV viral loads

用于诊断和监测 HIV 病毒载量的新型定量成本效益分析

• 批准号: 10320531
• 负责人: Karin Chumbimuni-Torres
• 金额: $ 7.14万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10320531
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Awareness; Bathing; Biological Assay; Biosensor; COVID-19 pandemic; Cessation of life; Counseling; Country; DNA; Data; Detection; Developing Countries; Diagnosis; Economics; Ensure; Epidemic; Equipment; Exclusion; Free Energy; Funding; Goals; HIV; HIV Genome; HIV Infections; HIV diagnosis; HIV-1; HIV-2; Hand; Hour; Human; Human immunodeficiency virus test; Hybrids; Individual; Infection; Joints; Methodology; Methods; Monitor; Nucleic Acid Amplification Tests; Nucleotides; Oligonucleotides; Patients; Performance; Polymerase Chain Reaction; Procedures; Protocols documentation; RNA; RNA amplification; Reaction; Research; Resources; Retroviridae; Reverse Transcription; Self-Sustained Sequence Replication; Signal Transduction; Stretching; Structure; T7 RNA polymerase; Techniques; Technology; Temperature; Time; United Nations; Validation; Variant; Viral Genome; Viral Load result; Viral load measurement; Virus; Visit; Water; amplification detection; base; cold temperature; cost effective; design; detection platform; falls; income disparities; infection rate; isothermal amplification; low and middle-income countries; new technology; novel; pandemic disease; point of care; point of care testing; point-of-care diagnostics; programs; promoter; rapid detection; treatment services; user-friendly; viral RNA; viral detection

PROJECT SUMMARY Since the start of the AIDS epidemic, according to the United Nations Joint Programme on HIV/AIDS (UNAIDS), a total of 75.7 million people have been infected with HIV, and of which 32.7 million have died due to AIDS- related illnesses. In 2016, UNAIDS defined several targets for HIV testing and treatment to be achieved by 2020 to end the AIDS epidemic by 2030. Such milestones include the reduction of the yearly infection rate to 500,000 new infections per year, the reduction of yearly AIDS-related deaths to 5000,000 deaths per year, and the 90- 90-90 goal, requiring that 90% of HIV-infected individuals be aware of their status, of which 90% are receiving treatment, and of which 90% have undetectable viral loads. We have now reached the goal year, and, based on data from 2019, we are not projected to meet any of these milestones. Although the targets were reached in a handful of countries, most low- and middle-income countries continue to fall behind the global goal in part due to decreases in funding and income disparities. To achieve these goals, extensive HIV testing is required, not only to diagnose new cases but in order to monitor the viral loads of those living with HIV. Therefore, there is an urgent need for a specific virus detection platform that can be used in low-resources settings. Here, we propose to develop a new technology for the specific, sensitive, and rapid detection of HIV-1 and HIV-2, using a universal recognition point-of-care testing platform. This unique platform integrates electrochemical detection of specific viral genome sequences using four-way junction (4WJ) probes following a modify isothermal amplification of the viral RNA fragments using Nucleic Acid Sequence Based Amplification (NASBA). The complete analysis (RNA amplification and detection) is expected to be executed in less than one hour, which is comparable with a regular visit to the doctor’s office. In the proposed research we will: 1) Optimized the modified-tag isothermal amplification using NASBA; 3) Design an electrochemical biosensor by selecting appropriate fragments of the amplicons to be interrogated by the 4WJ probes; 4) Optimize and characterize the biosensor for virus direct detection of amplicons; 5) Study variable parameters for single- and multiplex biosensor analysis; 6) Apply the developed biosensors for virus detection of commercially available reference material; and 7) Validate our methodology using a reference method (RT-qPCR). The proposed technology promises to be a user-friendly point-of-care testing at limited-resource settings for timely diagnosis of HIV infection and monitoring to help stopping the proliferation of AIDS infection by 2030.

项目摘要 根据联合国艾滋病毒/艾滋病联合计划（UNAIDS），自艾滋病流行病开始以来， 共有7570万人感染了艾滋病毒，其中3270万人因艾滋病而死亡 - 相关疾病。 2016年，UNAID定义了到2020年将实现HIV测试和治疗的几个目标 到2030年结束艾滋病流行。这些里程碑包括将年度感染率降低到500,000 每年的新感染，每年与AIDS相关的年死亡减少到每年5000,000人死亡，以及90- 90-90目标，要求90％的艾滋病毒感染者知道自己的状态，其中90％正在接受 治疗，其中90％的病毒负荷无法检测到。我们现在已经达到目标年，并基于 从2019年开始的数据，我们没有预计符合这些里程碑。尽管在 少数国家，大多数低收入国家和中等收入国家继续落后于全球目标 为了实现这些目标，需要进行广泛的艾滋病毒测试，而不是 仅是为了诊断新病例，但为了监测艾滋病毒感染者的病毒载荷。因此，有一个 迫切需要在低资源设置中使用的特定病毒检测平台。在这里，我们建议 使用通用 识别点测试平台。这个独特的平台集成了特定的电化学检测 在修改的等温扩增之后，使用四向连接（4WJ）问题的病毒基因组序列 使用基于核酸序列的扩增（NASBA）的病毒RNA片段。 完整的分析（RNA扩增和检测）预计将在不到一小时内执行 与定期访问医生办公室相当。在拟议的研究中，我们将：1）优化 使用NASBA修改标签等温扩增； 3）通过选择设计电化学生物传感器 4WJ探针询问的放大器的适当片段； 4）优化和表征 病毒直接检测扩增子的生物传感器； 5）研究单个和多个生物传感器的研究变量参数 分析; 6）应用开发的生物传感器以检测市售参考材料的病毒；和 7）使用参考方法（RT-QPCR）验证我们的方法。 该提议的技术有望在有限资源的设置上进行用户友好的护理测试 及时诊断HIV感染和监测，以帮助停止到2030年的艾滋病感染。