Surveying the Status of a Large Number of Mutations by A Single PEPD Assay

描述（由申请人提供）：本申请旨在开发PEPD技术，以调查大量过量野生型DNA中数百种DMA标记的突变状态，用于癌症筛查。具体来说，我们将开发一种粪便DMA检测方法用于结直肠癌（CRC）筛查。粪便DNA检测正在成为一种新的CRC筛查方法。然而，粪便DNA检测是一项技术挑战，因为没有单一的标记可以指示所有的crc，因此必须使用大量的标记来提高灵敏度。因此，现有的粪便DNA方法要么灵敏度差，要么成本高，不适合筛查。在第一阶段，我们证明了一种名为PEPD的新技术可以通过单次分析在超过正常DNA 1万倍的情况下调查许多突变，从而可以以一种敏感且经济有效的方式检测突变。据我们所知，这提供了第一个通过单一分析以如此高的灵敏度测量大量DNA标记状态的演示。PEPD方法简单，功能强大，可作为一种通用的突变检测技术平台。因此，该项目的成功将对突变分析和癌症筛查产生深远的影响。在II期开发的基于pepd的检测方法可以以经济有效的方式检测80-85%的crc。这项工作的成功对CRC筛查具有重要意义，因为它可以改变粪便DNA检测。最终，它允许医生通过简单地取一张粪便进行DNA分析来检测大多数结直肠癌，从而每年从结直肠癌中拯救数千人的生命。此外，该检测可用于监测结直肠癌患者和治疗后的风险评估。

DESCRIPTION (provided by applicant): This application is to develop the PEPD technology to survey the mutation status of hundreds of DMA markers in a vast excess of wild-type DNA for cancer screening. Specifically, we will develop a fecal DMA testing assay for colorectal cancer (CRC) screening. Fecal DNA testing is emerging as a novel method for CRC screening. However, fecal DNA testing is a technical challenge, as no single marker can indicate all CRCs and thus a large panel of markers must be utilized to increase sensitivity. As a result, existing fecal DNA methods are not suitable for screening because of either poor sensitivity or high-cost. In Phase I, we demonstrated that a novel technology called PEPD could survey a number of mutations in a 10,000 folds more excess of normal DNA by a single assay, making it possible to detect mutations in a sensitive and cost effective manner. To the best of our knowledge, this provides the first demonstration of surveying the status of a large number of DNA markers with such high sensitivity by a single assay. The PEPD method is simple, but very powerful and thus it can be a universal mutation detection technology platform. Hence, success of this project will have a profound impact on mutation analysis and thus cancer screening. The PEPD-based assay developed in Phase II can detect 80-85% of CRCs in a cost-effective manner. Success of this work is of great significance in CRC screening, as it can transform fecal DNA testing. Eventually, it allows physicians to detect the majority of CRC simply by taking a piece of stool for DNA analysis to save thousands of life each year from CRC. In addition, this assay can be used to monitor CRC patients and risk assessment after treatment.