COLOR DNA SEQUENCING APPARATUS FOR CLINICAL DIAGNOSIS

我们的目标是以p53基因为模型系统，研制一种适用于临床突变诊断的廉价自动化四色单泳道DNA测序仪。该机器的操作基于最近公开的荧光检测技术，该技术能够检测和分析低功率荧光信号。机器的主要特点：分析污染的DNA样品（野生和突变混合物的10%分辨率）; .高通量（高达500，000 bp/年）; .少量标记的DNA材料（10倍拉伸的ABI测序试剂盒）;机器的低成本在第一阶段，我们将与SUNY斯托尼布鲁克的电气工程系合作，开发一种数据传输速率为1.6 Mbit/s的超灵敏四色荧光检测系统，并与SUNY SB的病理学系合作，使用临床DNA材料测试该检测系统。在第二阶段，我们将开发一个完全基于半导体元件、集成技术和低功率照明源的机器的试验原型，并使用p53基因作为模型系统来验证机器。在第三阶段，我们将组织单毛细管机器的制造，并开发用于诊断应用的16通道测序仪。拟议的商业应用：拟议中的自动测序机将在遗传疾病（主要是癌症）的临床诊断中找到应用。其精确量化受污染DNA样本的独特能力将确保其在一个重要市场领域的主导地位。低成本，微型四色自动化机器与低消耗的标记DNA材料将是有吸引力的个人研究人员和医学诊断实验室。

Our goal is the development of an inexpensive automated 4-color single- lane DNA sequencing machine adapted for mutation diagnostics in clinical environment using the p53 gene as a model system. Operation of the machine is based on a recently disclosed multicolor fluorescent detection technique which enables the detection and analysis low-power fluorescence signals. Main features of the machine: . analysis of contaminated DNA samples (10% resolution of wild and mutated admixtures); . high throughput (up to 500,000 bp/year); . low amount of labeled DNA material (10 times stretched ABI sequencing kit); . low cost of the machine In Phase I in collaboration with the Department of Electrical Engineering, SUNY Stony Brook, we shall develop an ultra-sensitive 4-color fluorescent detection system with data transfer rate of 1.6 Mbit/s and, in collaboration with the Department of Pathology SUNY SB, test this detection system using clinical DNA material. In Phase II we shall develop a pilot prototype of the machine entirely based on semiconductor components, integrated technologies, and low-power illumination sources and validate the machine using the p53 gene as a model system. In Phase III we shall organize manufacturing of the single-capillary machine and develop a 16-lane sequencer for diagnostic applications. PROPOSED COMMERCIAL APPLICATIONS: The proposed automated sequencing machine will find applications in clinical diagnostics of genetic disease, primarily cancer. its unique capability to precisely quantify contaminated DNA samples will ensure a dominant position in an important market segment. The low cost, miniature 4-colored automated machine with low consumption of labeled DNA material will be attractive for individual researchers and medical diagnostic laboratories.