Low complexity, Instrument-free for CT/NG

描述(由申请人提供)：我们建议开发一种低成本、中等复杂性的分子检测方法，用于检测CT/NG，用于性病诊所。这项测试将使用手动操作的核酸提取设备、称为嗜热解旋酶依赖扩增(THDA)的等温放大技术、一次性核酸检测设备和可重复使用的读取器。我们的专利THDA使用解旋酶而不是加热来分离DNA链，但仍然依赖DNA聚合酶来扩增DNA，而不是聚合酶与其他酶的组合。这种简单的酶学方法使我们能够检测DNA扩增抑制剂，允许确认合法的产物被扩增，并对样本中的核酸进行定量。没有其他等温扩增技术保留了这些极具吸引力的特征。这两种检测的预计成本约为20美元。使用等温放大的优点是设备成本最低(即200美元的水浴和800美元的读取器)。我们建议将第一个项目年的重点放在进一步发展我们建议的检测系统上，通过提高反应速度，改进我们的原型样品准备模块，并演示使用带有最佳盒式磁带的自动阅读器的可行性。在第二年，我们将专注于正式的设计控制过程，并转移到我们建议的检测系统的制造。我们将制定一项临床研究计划，供机构IRBs审查，并制定一项前IDE，供食品和药物管理局(FDA)审查。在第三年和第四年，我们将进行一项多点临床研究，以获得向FDA提交510(K)计划的数据。我们预计在拟议的项目结束前获得FDA对CT/NG检测的批准。在第5年，我们将制定临床计划，并进行临床前研究，以评估获得CLIA豁免化验系统的可行性。

DESCRIPTION (provided by applicant): We propose to develop a low cost, moderate complexity molecular test for the detection of CT/NG intended for use at STD clinics. This test will use a manually operated nucleic acid extraction device, an isothermal amplification technology called thermophilic helicase dependent amplification (tHDA), a disposable nucleic acid detection device, and a reusable reader. Our proprietary tHDA uses helicases to separate DNA strands rather than heat but still relies on DNA polymerase to amplify DNA instead of combinations of polymerases with other enzymes. This simple enzymology allows us to detect DNA amplification inhibitors, allows for the confirmation that a legitimate product was amplified, and the quantitation of nucleic acids in a sample. No other isothermal amplification technology retains these highly attractive features. The projected cost for the two assays is ~$20. The advantage of using isothermal amplification is that equipment costs will be minimal (i.e., $200 water bath, and a ~$800 reader). We propose to focus our first project year on further evolving our proposed assay system by improving our reaction speed, improving our prototype sample preparation module, and demonstrating the feasibility of using an automated reader with the BESt cassette. In year 2, we will focus on the formal design control process, and transfer to manufacturing of our proposed assay system. We will develop a clinical study plan for review by institutional IRBs, and a pre-IDE for review by the Food and Drug Administration (FDA). In years 3 and 4 we will perform a multi-site clinical study to obtain data for the submission of a 510(k) to the FDA. We expect to obtain FDA clearance for the CT/NG assays by the end of the proposed project. In year 5, we will develop a clinical plan, and perform a pre-clinical study to evaluate the feasibility of obtaining CLIA-waiver for the assay system.