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）：在该I期项目中，我们提出研究一种称为Zip核酸（ZNA”）的新型引物化学和探针检测系统。ZNA是与许多阳离子精胺部分缀合的寡核苷酸，所述阳离子精胺部分增强引物和探针在核酸靶标附近的有效浓度。 据报道，该特性可提高RT-PCR的速度和灵敏度（Moreau et al. 2009）。 ZNA与Taqman检测格式兼容（巴黎等人，2010）。 我们预计这项检测技术将进一步加速我们的RNA检测分析并提高我们的准确性。 我们将比较ZNA与LNA DNA双标记（Tong et al. 2008，Li et al. 2010）和CPT探针的性能。 在研究结束时，我们将完成Isopropyl HIV-1定量检测试剂盒商业放行所需的分析研究阶段。 我们的第一阶段研究计划包括四个目标： 1.设计并测试针对HIV-1所有亚型的ZNA引物， 2.设计和测试ZNA taqman和ZNA循环探针用于HIV检测， 3.开发从干血斑（DBS）和干血浆中提取RNA的简单工作流程 点（DPS），以及 4.使用ZNA技术结合以下技术测试检测的灵敏度和特异性： 一组HIV-1分离株中的异丙肾上腺素。 在第一阶段结束时，我们将准备确定最佳的探针技术，以开发一种用于美国和国外商业销售的商业Isoprotein HIV-1定量检测。 我们还将明确指出最适合HDA病毒载量检测的样本提取方法的类型。 在第二阶段，我们将为多中心临床研究计划开发一个预IDE，以寻求FDA批准在美国销售。 我们还将探索在世界其他地区的商业发行。