Biosensor Assays for Diagnosis of Chronic HCV Infection

用于诊断慢性 HCV 感染的生物传感器测定

• 批准号: 8646475
• 负责人: ROBERT W HENKENS
• 金额: $ 13.73万
• 依托单位: ALDERON BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-05 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8646475
• 关键词: Academic Medical Centers; American; Bedside Testings; Biological Assay; Biosensor; Blood capillaries; Caring; Centers for Disease Control and Prevention (U.S.); Characteristics; Chronic Hepatitis C; Clinic; Clinical; Communities; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Direct Costs; Disease; Enzymes; Epidemic; FDA approved; Facilities and Administrative Costs; HIV; Health; Healthcare; Hepatitis C virus; Human; Individual; Infection; Intervention; Label; Left; Legal patent; Measurement; Measures; Methods; Molecular; Monitor; Neighborhoods; Optics; Patients; Performance; Phase; Physiologic pulse; Plasma; Positioning Attribute; Primer Extension; Printing; Procedures; Qualifying; RNA; RNA Viruses; Reader; Research; Research Personnel; Resources; Sampling; Serum; Signal Transduction; Small Business Innovation Research Grant; Specimen; System; Techniques; Technology; Testing; Treatment Protocols; Universities; Viral; Viral hepatitis; base; capillary; cost; design; disorder prevention; electrical measurement; experience; innovation; member; novel; novel strategies; pathogen; point of care; public health relevance; research and development; response; success; technology development; tool; viral RNA; viral detection; years of life lost

DESCRIPTION (provided by applicant): This Phase I SBIR application is submitted in response to the call by DHHS for new tests for qualitative measurement of HCV RNA for diagnosis of chronic HCV infection in patients. The need is for HCV RNA assays that are as reliable but less expensive and resource-demanding than current molecular assays. The new test proposed for research and development consists of rapid sample isolation, innovative signal amplification, and detection with a new proprietary Alderon biosensor. The innovation in this research lies in the creation of a new, inexpensive, low-cost, assay for HCV RNA by development of integrated RNA isolation and advanced electrochemical (eSystem) detection, whose feasibility and performance characteristics will be investigated in research with three specific aims. In Aim 1, we will seek to demonstrate a simple mode of making the viral RNA in biofluid samples available for our downstream eSystem assays. In Aim 2, we will investigate innovative HCV RNA detection using a patented signal amplification method (PESA) and a new capillary flow design for advanced, low-cost biosensors that generate target-dependent electrical currents that are measured with an eSystem reader/monitor developed by us. The assays will employ direct detection of target-specific enzyme labels for high sensitivity, without the wash steps usually required for enzyme-enhanced assays. Our low-cost readers will deliver controlled electrical pulses and monitor currents generated by the target-specific enzyme labels. The milestone for Aim 2 will be to determine whether this new mode of HCV RNA detection has a sensitive electrical response with accurate detection down to 500 IU/mL. The objective is a biosensor system with performance equal to that of existing FDA-approved, branched-probe (bProbe) assays, yet practical for wide spread use at point of care. In these Aim 2 studies, we will use Armored HCV RNA standards and mock clinical specimens consisting of the hepatitis C virus spiked into serum and plasma samples. In Aim 3, we will validate the Alderon HCV RNA Qualitative eSystem Assay with clinical specimens. Accomplishing these aims will show the feasibility of developing new eSystem qualitative HCV RNA tests that are as clinically useful and reliable as commercially available, FDA-approved tests for confirmatory testing of chronic HCV infections but at much lower cost and suitable for use in resource-limited communities. We anticipate that this new approach to qualitative HCV RNA testing will find use both as a clinical diagnostic and as a valuable research tool for the study of HCV chronic infections. The proposed eSystem technology to be developed in this project will have a high impact, as it brings lower-cost HCV testing to many more infected individuals. Additional impacts of project success will result from use of the basic technology to develop new tests for other RNA viruses, such as HIV.

描述（由申请人提供）：本I期SBIR申请是应DHHS要求提交用于诊断慢性HCV感染的HCV RNA定性测量新测试的要求而提交的。需要的是与目前的分子检测方法一样可靠但更便宜和资源需求更少的HCV RNA检测方法。新测试提出的研究和开发包括快速样品分离，创新的信号放大和检测与新的专有Alderon生物传感器。本研究的创新之处在于通过开发集成RNA分离和先进电化学（essystem）检测技术，创造了一种新的、廉价的、低成本的HCV RNA检测方法，其可行性和性能特征将在三个具体目标下进行研究。在目标1中，我们将寻求证明一种简单的模式，使生物流体样品中的病毒RNA可用于我们的下游系统分析。在目标2中，我们将研究创新的HCV RNA检测，使用专利的信号放大方法（PESA）和一种新的毛细流设计，用于先进的低成本生物传感器，该传感器产生依赖靶标的电流，并使用我们开发的eSystem读取器/监视器进行测量。该分析将采用直接检测目标特异性酶标记的高灵敏度，而不需要酶增强分析通常需要的洗涤步骤。我们的低成本阅读器将提供受控的电脉冲和监测由目标特异性酶标签产生的电流。Aim 2的里程碑将是确定这种新的HCV RNA检测模式是否具有敏感的电反应，准确检测低至500 IU/mL。目标是一种生物传感器系统，具有与现有fda批准的分支探针（bProbe）检测相同的性能，但可在护理点广泛使用。在这些Aim 2研究中，我们将使用装甲HCV RNA标准和模拟临床标本，包括加入血清和血浆样本的丙型肝炎病毒。在目标3中，我们将用临床标本验证Alderon HCV RNA定性系统检测。实现这些目标将表明开发新的essystem定性HCV RNA测试的可行性，这些测试在临床有用和可靠程度上与市售的、fda批准的慢性HCV感染确证测试一样，但成本要低得多，适合在资源有限的社区使用。我们期待这种定性HCV RNA检测的新方法将作为临床诊断和HCV慢性感染研究的有价值的研究工具。该项目拟开发的eSystem技术将产生重大影响，因为它将为更多的感染者带来成本更低的丙型肝炎病毒检测。项目成功的其他影响将来自使用基本技术开发其他RNA病毒（如艾滋病毒）的新检测方法。