Twist-Biosensor: Novel single molecule microarray technology for DNA genotyping

描述(申请人提供)：人类基因组信息的可获得性及其变异性使研究人员能够确定导致患者对治疗的不同反应的基因变异。1-3为了将个性化医学的好处提供给临床实践，有必要拥有能够确定多个遗传标记的准确基因分型技术。现有的食品和药物管理局(FDA)批准的药物遗传学测试需要很长的处理时间或具有有限的多功能性。在这里，我们提出了基于Twist生物传感器技术的快速和多重药物遗传检测的发展。扭曲生物传感器是基于单分子DNA杂交和超螺旋技术的新型寡核苷酸检测微阵列技术。在这种技术中，杂交靶标受到破坏性扭转应力，这允许使用简单的设备以极低的背景噪声快速检测多个单核苷酸多态(SNPs)。我们的总体目标是展示基于Twist生物传感器技术的设备可以在从样品到结果的三个小时内在单个芯片上对几个SNP进行基因分型，从而使患者更容易获得个性化药物。我们的方法将是建立一个原型设备并对其进行表征，该设备能够对CYP2D6基因的三个SNPs进行基因分型，该基因是药物标签上发现的最普遍的基因组生物标记物。在美国，大约13%的患者接受由CYP2D6酶代谢的处方药。4目标1专注于快速检测单个CYP2D6突变。目的2研究快速检测小样本中多个CYP2D6单核苷酸多态的方法。

DESCRIPTION (provided by applicant): The availability of human genomic information and its variability is allowing researchers to identify the genetic variations responsible for patients' different responses to treatment.1-3 In order to deliver the benefits of personalized medicine to the clinical practice, it is necessary to have accurate genotyping techniques capable of determining multiple genetic markers. The existing Food and Drug Administration (FDA)-approved pharmacogenetics tests require long processing times or have limited multiplexability. Here, we propose the development of rapid and multiplex pharmacogenetic tests based on Twist-Biosensor technology. Twist-Biosensor is a novel microarray technology for oligonucleotide detection based on single molecule DNA hybridization and supercoiling. In this technique, the hybridized targets are subjected to disrupting torsional stress which allows for rapid detection of multiple Single Nucleotide Polymorphisms (SNPs) with extremely low background noise using a simple device. Our overall aim is to demonstrate that devices based on the Twist-Biosensor technology can make personalized medicine more readily available to patients by genotyping several SNPs in a single chip within three hours from sample to results. Our approach will be to build and characterize a prototype device able to genotype three SNPs of the CYP2D6 gene, which is the most prevalent genomic biomarker found on drug labels. About 13% of patients in the U.S. receive prescription drugs metabolized by the CYP2D6 enzyme.4 Aim 1 focuses on fast detection of a single CYP2D6 mutation. Aim 2 focuses on fast multiple CYP2D6 SNP detection in small blood samples.