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、中等复杂度的分子检测方法，用于检测性病诊所使用的CT/NG。该检测将使用手动操作的核酸提取装置、称为嗜热解旋酶依赖性扩增（tHDA）的等温扩增技术、一次性核酸检测装置和可重复使用的读数器。我们专有的tHDA使用解旋酶而不是加热来分离DNA链，但仍然依赖于DNA聚合酶来扩增DNA，而不是聚合酶与其他酶的组合。这种简单的酶学允许我们检测DNA扩增抑制剂，允许确认合法的产物被扩增，并定量样品中的核酸。没有其他等温扩增技术保留这些高度吸引人的特征。两种检测的预计成本约为20美元。使用等温扩增的优点是设备成本将是最小的（即，200美元的水浴和800美元的阅读器）。我们建议将第一个项目年的重点放在通过提高反应速度、改进原型样品制备模块以及证明使用自动化阅读器和BEST试剂盒的可行性来进一步发展我们提出的检测系统。在第2年，我们将专注于正式设计控制过程，并转移到我们拟定的检测系统的生产。我们将制定临床研究计划供机构IRB审查，并制定预IDE供食品药品监督管理局（FDA）审查。在第3年和第4年，我们将进行多中心临床研究，以获得向FDA提交510（k）的数据。我们希望在拟定项目结束时获得CT/NG检测试剂盒的FDA批准。在第5年，我们将制定临床计划，并进行临床前研究，以评估获得检测系统CLIA豁免的可行性。

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.