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

项目描述（由申请人提供）：在本一期项目中，我们拟研究一种新型的引物化学和探针检测系统，称为Zip核酸（ZNA）。ZNAs是与一些阳离子精胺基团结合的寡核苷酸，可以提高核酸靶点附近引物和探针的有效浓度。据报道，这一特性可提高RT-PCR的速度和灵敏度（Moreau et al. 2009）。zna与Taqman检测格式兼容（Paris et al. 2010）。我们期望这项检测技术将进一步加快我们的RNA检测分析，并提高我们的准确性。我们将比较ZNA与LNA DNA双标记探针（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IsoAmp HIV-1定量测定的商业发布所需的分析研究阶段。我们一期的研究计划包括4个目标：1。设计和测试针对所有hiv亚型的ZNA引物1,2。设计和测试ZNA taqman和ZNA循环探针用于HIV检测。3 .建立从干血斑（DBS）和干血浆斑（DPS）中提取RNA的简单工作流程；在HIV-1分离株中，使用ZNA技术联合IsoAmp测试检测方法的敏感性和特异性。在I期结束时，我们将准备确定最佳探针技术，以开发商业化IsoAmp HIV-1定量分析，以便在美国和国外进行商业分销。我们也将有一个明确的指示类型的样品提取方法，最适合HDA病毒载量测试。在第二阶段，我们将开发一个多地点临床研究计划的预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。

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.