Single molecule microarrays for the detection of mutant DNA in body fluids

用于检测体液中突变 DNA 的单分子微阵列

• 批准号: 8625158
• 负责人: Alfredo Andres Celedon
• 金额: $ 22.89万
• 依托单位: SCANOGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-19 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8625158
• 关键词: Alleles; Biological Assay; Biological Markers; Biosensor; Blood; Body Fluids; Cancer Diagnostics; Cancer Patient; Cell Proliferation; Cells; Complex; DNA; DNA Fingerprinting; DNA amplification; Detection; Development; Diagnostic Procedure; Early Diagnosis; Electronics; Feces; Gene Mutation; Hour; Hybrids; Individual; KRAS2 Gene Mutation; KRAS2 gene; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Methods; Molecular; Molecular Diagnostic Techniques; Mutate; Mutation; Mutation Detection; Nucleotides; Oligonucleotides; Oncogenes; Plasma; Point Mutation; Procedures; Property; Resolution; Sampling; Screening for cancer; Staging; Stress; Techniques; Technology; Testing; Time; Tumor Cell Line; Urine; base; cancer microarray; cost effective; design; improved; multiplex detection; mutant; neoplastic cell; new technology; novel; novel strategies; prevent; public health relevance; rapid detection; screening; sensor; single molecule; synthetic construct; tool

DESCRIPTION (provided by applicant): The formation of malignant tumors is associated with accumulation of genetic mutations that enhance cell proliferation. Detecting these mutations in body fluids can be used for cancer management and early cancer detection. However, current cancer diagnostic methods are not well suited for the detection of mutant DNA in body fluids which have prevented the development of effective, cost-efficient cancer screening strategies. Here, we propose the development of microarrays based on Twist-Biosensor technology to dramatically improve and simplify detection of biomarkers in body fluids for cancer screening. Twist-Biosensor is a novel microarray technique in which hybridization is detected with single molecule resolution. In addition, Twist-Biosensor applies disrupting torsional stress to DNA hybrids, a novel strategy uniquely designed to give microarrays high point mutation selectivity. Therefore, the technique is ideal for detection of mutated DNA at low concentration in the presence of a large proportion of wild-type DNA. The overall aim of this application is to demonstrate the capabilities of Twist-Biosensor microarrays for cancer screening. We will develop assays to detect mutations of the KRAS gene and test them with plasma and urine samples from pancreatic cancer patients. Each of the proposed aims of this application focuses on establishing two complementary properties of the microarrays: Detection of multiple cancer mutations (Aim 1) and ultra-sensitive detection of mutated DNA (Aim 2). In addition, both aims will demonstrate that Twist-Biosensor microarrays are faster and require fewer steps than alternative diagnostic techniques.

描述（由申请人提供）：恶性肿瘤的形成与增强细胞增殖的基因突变的积累有关。检测体液中的这些突变可用于癌症管理和早期癌症检测。然而，目前的癌症诊断方法不太适合检测体液中的突变DNA，这阻碍了有效的、具有成本效益的癌症筛查策略的发展。在这里，我们提出了基于Twist-Biosensor技术的微阵列的发展，以显着提高和简化检测体液中的生物标志物的癌症筛查。Twist-Biosensor是一种新型的微阵列技术，它可以在单分子水平上检测杂交。此外，Twist-Biosensor将破坏性扭转应力应用于DNA杂交，这是一种独特设计的新策略，可使微阵列具有高点突变选择性。因此，该技术对于在存在大比例野生型DNA的情况下检测低浓度的突变DNA是理想的。本申请的总体目标是证明Twist-Biosensor微阵列用于癌症筛查的能力。我们将开发检测KRAS基因突变的方法，并使用胰腺癌患者的血浆和尿液样本进行检测。本申请的每个目标都集中在建立微阵列的两个互补特性：检测多种癌症突变（目标1）和超灵敏检测突变DNA（目标2）。此外，这两个目标都将证明Twist-Biosensor微阵列比其他诊断技术更快，需要的步骤更少。