Electrochemical RT Activity Assay for Measuring HIV Load

用于测量 HIV 负荷的电化学 RT 活性测定

• 批准号: 8015016
• 负责人: ROBERT W HENKENS
• 金额: $ 47.06万
• 依托单位: ALDERON BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8015016
• 关键词: Acquired Immunodeficiency Syndrome; Address; Affect; Biological Assay; Caring; Child; Clinical; Complex; Consult; Detection; Development; Development Plans; Diagnosis; Diagnostic; Diagnostic tests; Diamond; Disasters; Disease; Engineering; Environment; Enzymes; FDA approved; Geographic Locations; Goals; HIV; HIV Seropositivity; HIV-1; HIV-2; Health; Healthcare; Individual; Laboratories; Laboratory Research; Lead; Letters; Life; Measurement; Measures; Methods; Molecular Biology; Monitor; National Institute of Allergy and Infectious Disease; Optics; Patients; Performance; Pharmaceutical Preparations; Phase; Phase III Clinical Trials; Play; Poverty; Predisposition; Primary Health Care; Procedures; Product Approvals; Protocols documentation; RNA-Directed DNA Polymerase; Reagent; Relative (related person); Research; Research Priority; Research Project Grants; Resources; Role; Sales; Sampling; Series; Small Business Innovation Research Grant; Staging; System; Technology; Testing; Therapeutic; Therapeutic Intervention; Time; TimeLine; Validation; Viral; Viral Load result; Viral Physiology; Viral load measurement; Work; World Health Organization; base; commercialization; cost; health care quality; improved; innovation; instrument; instrumentation; man; novel diagnostics; point of care; prevent; product development; research and development; research facility; sensor; success; tool; viral RNA; virology

DESCRIPTION (provided by applicant): To address the urgent need for better treatment and diagnostic testing of HIV infected people in resource-limited settings, we propose in this Phase II SBIR to further develop an electrochemical testing system (eSystem) developed by Alderon Biosciences, Inc. for determinations of HIV viral load in clinical samples. Alderon's long-term goal is to produce, validate, and gain FDA approval for a facile and affordable (< $10) HIV viral load test, sensitive to below 500 copies/ml, that gives same-day results using an inexpensive and portable instrument. Currently available FDA-approved HIV viral load tests require expensive instrumentation, utilize complex molecular biology procedures, and can cost > $100 per test. Our hypothesis is that Alderon can develop a far more facile and affordable system for accurate viral load measurements and drug susceptibility testing that will enable viral load testing to be done in resource-limited point-of-care (POC) environments. The innovation in the new HIV viral load tests proposed by Alderon lies in the use of advanced electrochemical detection methods for quantifying the activity of reverse transcriptase (RT) in clinical samples. The viral RT enzyme plays an essential role in viral replication, regardless of HIV subtype. Because the enzyme's presence is highly correlated with that of the viral RNA, RT activity assays are a recognized (but not FDA-approved) alternative method for monitoring viral load. Unlike PCR-based assays that have problems quantifying the often-changing RNAs of some HIV subtypes, RT activity tests work equally well for HIV-1 (regardless of subtype) and HIV-2, can be done without highly skilled technicians, and are less subject to problems of contamination. Alderon's Phase I work has shown feasibility of an electrochemical approach to quantification of viral RT activity, with the potential for greatly improved performance (time and cost per test) relative to optical assays. Alderon's proposed 3 year Phase II SBIR project has 4 Specific Aims that will lead to eSystem improvements and accurate detection of HIV load in facile and low-cost same-day tests. Aims 1 and 2 focus on improvements in the instrument and sensors that will make the viral load assays more affordable and simpler for use in resource-limited settings. Aim 3 improvements of reagents and assay protocols will further reduce assay time and complexity. Aim 4 validation of eSystem performance in viral load tests with a wide range of samples will set the stage for FDA approval and product commercialization. Collaborative work with the UNC Center for AIDS Research will establish the correlation between results of eSystem assays and viral load results obtained by alternative FDA- approved methods. Product commercialization will be facilitated by collaborative work with Magellan Biosciences, whose engineering expertise will aid in the development of reliable and robust instruments and viral load test kit components that can be produced in facilities certified for good manufacturing practices. PUBLIC HEALTH RELEVANCE: An estimated 33.2 million people are now living with HIV disease worldwide and in 2007 it claimed the lives 2.1 million people, including 330,000 children (World Health Organization). To address the urgent need for better treatment and diagnostic testing of HIV infected people, Alderon Biosciences, Inc. proposes to further develop a low-cost and easy-to-use electrochemical testing system (eSystem) for determinations of HIV viral load in clinical samples. Alderon's approach will make it possible to perform viral load testing in primary care or point-of-care (POC) environments and aid in monitoring and treating HIV positive patients in resource-limited environments.

描述（由申请人提供）：为了解决在资源有限的环境下对HIV感染者更好的治疗和诊断测试的迫切需求，我们在这个二期SBIR中提议进一步开发由Alderon Biosciences， Inc.开发的电化学测试系统（essystem），用于测定临床样品中的HIV病毒载量。Alderon的长期目标是生产、验证并获得FDA批准的一种简便且价格合理（< 10美元）的HIV病毒载量测试，对低于500拷贝/ml的病毒载量敏感，使用廉价的便携式仪器可在当天获得结果。目前，fda批准的HIV病毒载量测试需要昂贵的仪器，使用复杂的分子生物学程序，每次测试的成本可能高达100美元。我们的假设是，Alderon可以开发一种更方便和负担得起的系统，用于准确的病毒载量测量和药物敏感性测试，这将使病毒载量测试能够在资源有限的护理点（POC）环境中进行。Alderon提出的新型HIV病毒载量检测的创新之处在于使用先进的电化学检测方法来定量临床样品中逆转录酶（RT）的活性。无论HIV亚型如何，病毒RT酶在病毒复制中起着至关重要的作用。由于酶的存在与病毒RNA的存在高度相关，RT活性测定是一种公认的（但未经fda批准的）监测病毒载量的替代方法。与基于聚合酶链反应的分析不同，RT活性测试对HIV-1（无论何种亚型）和HIV-2同样有效，可以在没有高技能技术人员的情况下完成，并且较少受到污染问题的影响。Alderon的第一阶段工作已经证明了电化学方法量化病毒RT活性的可行性，与光学分析相比，有可能大大提高性能（每次测试的时间和成本）。Alderon提出的为期3年的二期SBIR项目有4个具体目标，这些目标将导致系统改进，并通过方便和低成本的当日检测准确检测艾滋病毒载量。目标1和目标2的重点是改进仪器和传感器，使病毒载量测定在资源有限的情况下使用起来更经济、更简单。目的3试剂和分析方案的改进将进一步减少分析时间和复杂性。Aim 4：在大范围样品中验证系统在病毒载量测试中的性能将为FDA批准和产品商业化奠定基础。与UNC艾滋病研究中心的合作将建立eSystem检测结果与通过FDA批准的替代方法获得的病毒载量结果之间的相关性。与麦哲伦生物科学公司的合作将促进产品商业化，麦哲伦生物科学公司的工程专业知识将有助于开发可靠和强大的仪器和病毒载量测试试剂盒组件，这些组件可以在经过良好生产规范认证的设施中生产。公共卫生相关性：目前全世界估计有3 320万人感染艾滋病毒，2007年有210万人死于艾滋病毒，其中包括33万名儿童（世界卫生组织）。为了满足对HIV感染者更好的治疗和诊断测试的迫切需求，Alderon Biosciences, Inc.提出进一步开发一种低成本、易于使用的电化学测试系统（essystem），用于测定临床样品中的HIV病毒载量。Alderon的方法将使在初级保健或护理点（POC）环境中进行病毒载量检测成为可能，并有助于在资源有限的环境中监测和治疗艾滋病毒阳性患者。