Rapid Assay for the l1307K Mutation of the APC Gene

描述(由申请人提供)：本提案描述了一种基于金纳米颗粒探针的简单、快速且经济实惠的单核苷酸多态(SNP)检测平台的开发。在第一阶段，高选择性的金纳米探针可用于快速、可靠地对含有I1307K突变的APC基因序列进行SNP基因分型。当探针结合到DNA靶标上时产生的比色变化通过在膜上斑点来记录，并使用简单的平板扫描仪或肉眼可视化。在第二阶段，纳米球公司寻求将这种新的分析方法扩展到对SNP的大规模分析。这将通过开发一种新的斑点方法和简单的比色读取器/软件包来实现，该方法能够在最少的用户干预下快速发现和分析多个PCR样本，以确定是否存在特定的DNA序列。然后，这些进展将被用来迅速将分析扩展到与结直肠癌、乳腺癌和心血管疾病相关的其他SNP和突变(例如插入和缺失)。这项技术的发展最终将填补一个既快速又简单的具有成本效益和可靠的SNP检测平台的空白。

DESCRIPTION (provided by applicant): This proposal describes the development of a simple, rapid and cost effective 'spot and read' detection platform for single nucleotide polymorphisms (SNP's) based on gold nanoparticle probes. In phase I, it was demonstrated that the highly selective gold nanoparticle probes could be used for rapid and reliable SNP genotyping of PCR-amplified APC gene sequences containing the I1307K mutation. The colorimetric change generated by the probes when bound to DNA target was recorded by spotting onto a membrane and visualized using a simple flatbed scanner or the naked eye. In phase II, Nanosphere seeks to extend this new assay to large scale analysis of SNP's. This will be accomplished by developing a new spotting methodology and simple colorimetric reader/software package that enables multiple PCR samples to be spotted rapidly and analyzed for the presence of specific DNA sequences with minimal user intervention. These developments will then be used to rapidly extend the assay to other SNP's and mutations (e.g. insertions and deletions) related to colorectal cancer, breast cancer, and cardiovascular disease. The development of this technology will ultimately fill the void of a cost effective and reliable SNP detection platform that is both rapid and simple.