RAPID ASSAY FOR THE I1307K MUTATION OF THE APC GENE

APC 基因 I1307K 突变的快速检测

• 批准号: 6074583
• 负责人: JAMES J. STORHOFF
• 金额: $ 10万
• 依托单位: NANOSPHERE, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-15 至 2001-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6074583
• 关键词: colloids; colorectal neoplasms; colorimetry; gene mutation; genetic markers; neoplasm /cancer genetics; nucleic acid hybridization; oligonucleotides; technology /technique development

The innovative use of oligonucleotide colloidal gold nanoparticle conjugates is proposed as a simple, rapid and inexpensive method for the detection of Polymerase Chain Reaction (PCR) amplified nucleic acids of the I1307K mutation of the APC gene. The APC gene is known to play an important role in colorectal cancer (CRC). To date, only sequencing, which requires two days of processing and is very expensive, has been utilized for screening. Preliminary examination of this new class of hybridization probes in conjunction with single stranded synthetic oligonucleotide targets already has demonstrated the potential of this approach for the detection of a model synthetic target sequence. Specifically, single base imperfections, regardless of position along the target strand, can be differentiated easily from a perfect target sequence using this novel colorimetric technique. This detection strategy is based on the aggregation of colloidal gold particles which leads to changes in the particle surface plasmon band. We propose to further develop this method in order to generate a rugged and routine technique for analysis of the PCR products of the I307K APC gene mutation. This would provide the first molecular probe based assay for screening of this important cancer gene. PROPOSED COMMERCIAL APPLICATIONS: A faster, simpler assay for screening mutations of the APC gene would facilitate early diagnosis of this deadly disease.

创新性地利用寡核苷酸胶体金纳米颗粒偶联物作为一种简单、快速、廉价的检测APC基因I1307K突变的聚合酶链反应（PCR）扩增核酸的方法。已知APC基因在结直肠癌（CRC）中起重要作用。迄今为止，只有测序被用于筛选，这需要两天的处理时间，而且非常昂贵。对这种新型杂交探针与单链合成寡核苷酸靶标结合的初步研究已经证明了这种方法检测模型合成靶标序列的潜力。具体来说，单碱基缺陷，无论沿着目标链的位置如何，都可以使用这种新的比色技术轻松地与完美的目标序列区分开来。这种检测策略是基于胶体金粒子的聚集导致粒子表面等离子体带的变化。我们建议进一步发展这一方法，以产生一种可靠和常规的技术来分析I307K APC基因突变的PCR产物。这将为筛选这一重要的癌症基因提供第一个基于分子探针的检测方法。建议的商业应用：一种更快、更简单的APC基因突变筛选方法将有助于这种致命疾病的早期诊断。