Rapid Assay for the l1307K Mutation of the APC Gene

描述（由申请人提供）：该提案描述了基于金纳米颗粒探针的单核苷酸多态性（SNP）的简单，快速和经济高效的“斑点和读取”检测平台的开发。在第一阶段，高选择性金纳米颗粒探针可用于快速可靠的pcr扩增APC基因序列的SNP基因分型，其中包含I1307K突变。探针与DNA目标结合时产生的比色变化通过点射到膜上记录下来，并使用简单的平板扫描仪或肉眼可视化。在第二阶段，Nanosphere试图将这种新方法扩展到SNP的大规模分析。这将通过开发一种新的定位方法和简单的比色阅读器/软件包来实现，该软件包使多个PCR样品能够快速定位并分析特定DNA序列的存在，而用户干预最少。随后，这些进展将用于快速将该检测扩展到与结直肠癌、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）。这项技术的发展将最终填补成本效益高、可靠、快速、简单的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.