Low-cost, rapid quantitative Isothermal Assay for HIV RNA using ZNA

使用 ZNA 对 HIV RNA 进行低成本、快速定量等温测定

• 批准号: 8426086
• 负责人: Huimin Kong
• 金额: $ 30万
• 依托单位: BIOHELIX CORPORATION
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-15 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8426086
• 关键词: Adherence; Affinity; Anti-Retroviral Agents; Back; Binding; Biological Assay; Blood; Chemistry; Clinical; Country; DNA; DNA-Directed DNA Polymerase; Detection; Dose; Economically Deprived Population; Economics; Enzymes; Equipment; Europe; Failure; Fluorescence; Goals; Human; Human Resources; Immune; Label; Laboratory Personnel; Licensing; Logistics; Measurement; Methods; Minor; Molecular; Molecular Conformation; Monitor; Multi-Institutional Clinical Trial; National Institute of Allergy and Infectious Disease; Nucleic Acid Probes; Nucleic Acids; Oligonucleotides; Patients; Performance; Pharmaceutical Preparations; Phase; Plasma; Polymerase Chain Reaction; Problem Solving; Property; RNA; Reporting; Research; Resources; Rest; Reverse Transcription; Risk; Sales; Sampling; Sensitivity and Specificity; Signal Transduction; Small Business Innovation Research Grant; Sodium Chloride; Specimen; Speed; Spermine; Spottings; Staging; System; Taq Polymerase; Technology; Temperature; Testing; Therapeutic; Time; Training; United States; Variant; Viral Load result; Viral load measurement; Virus; Work; base; cost; cost effective; design; health care delivery; helicase; inhibitor/antagonist; instrument; instrumentation; internal control; locked nucleic acid; melting; mortality; novel; phase 2 study; response; standard of care; viral RNA

DESCRIPTION (provided by applicant): In this Phase I project, we propose to investigate a novel primer chemistry and probe detection system called the Zip nucleic acids (ZNA"). ZNAs are oligonucleotides conjugated to a number of cationic spermine moieties that enhance the effective concentration of primers and probes near nucleic acid targets. This property has been reported to enhance the speed and sensitivity of RT-PCR (Moreau et al. 2009). ZNAs are compatible with Taqman detection formats (Paris et al. 2010). We expect this detection technology will further accelerate our RNA detection assays as well as increase our accuracy. We will compare the performance of ZNA to LNA DNA dual labeled (Tong et al. 2008, Li et al. 2010) and CPT probes. By the end of the study we will have completed the analytical study stage required for the commercial release of an IsoAmp HIV-1 quantitative assay. Our phase I research plan includes 4 aims: 1. Design and test ZNA primers targeting all subtypes of HIV-1, 2. Design and test with both ZNA taqman and ZNA cycling probes for the HIV assay, 3. Develop a simple work flow for extraction of RNA from dry blood spots (DBS) and dry plasma spots (DPS), and 4. Test the sensitivity and specificity of assays using the ZNA technology in combination with IsoAmp in a panel of HIV-1 isolates. At the conclusion of Phase I, we will be ready to identify the best probe technology to develop a commercial IsoAmp HIV-1 quantitative assay for commercial distribution in the US and abroad. We will also have a clear indication of the type of sample extraction method that best suits HDA viral load testing. In Phase II, we would develop a pre-IDE for a multi-site clinical study plan to seek FDA approval for sale in the US. We would also explore commercial release in the rest of the World.

描述（由申请人提供）：在此I阶段项目中，我们建议研究一种称为Zip核酸（ZNA”）的新型底漆化学和探针检测系​​统。ZnAS是寡核苷酸寡核苷酸，与许多阳离子精子部分结合在一起，以增强有效的有效性据报道，核酸靶标在核酸方面的浓度增强了RT-PCR的速度和敏感性（Moreau etal。2009）这种检测技术将进一步加速我们的RNA检测测定法，并提高我们的精度。我们将完成Isoamp HIV-1定量测定的商业释放所需的分析研究阶段。使用ZNA Taqman和ZNA循环探针进行设计和测试，用于HIV分析，3。开发一个简单的工作流，用于从干血点（DBS）和干等离子体斑点（DPS）（DPS）中提取RNA和4。测试敏感性和特异性使用ZNA技术与HIV-1分离株小组组合的ISOAMP结合使用的测定。 在第一阶段结束时，我们将准备确定最佳的探测技术，以开发用于美国和国外商业分销的商业ISOAMP HIV-1定量测定法。 我们还将清楚地表明最适合HDA病毒载荷测试的样品提取方法的类型。 在第二阶段，我们将为多站点临床研究计划制定一个预ID，以寻求在美国出售的FDA批准。 我们还将探索世界其他地区的商业发布。