用于基因突变和SNP分析的高分辨熔解曲线分析仪研制

项目名称： 用于基因突变和SNP分析的高分辨熔解曲线分析仪研制

项目编号： No.61227018

项目类型： 专项基金项目

立项/批准年度： 2013

项目学科： 无线电电子学、电信技术

项目作者： 彭年才

作者单位： 西安交通大学

项目金额： 290万元

中文摘要： 高分辨熔解曲线分析（HRM）是基因突变检测和SNP分析的新方法，为解决科学研究缺乏HRM分析仪的瓶颈，项目通过分析最近邻热力学模型缺陷，提出运用free-form特征提取发现复杂DNA熔解过程中物理规律的新思想，并研制高通量的专用HRM分析仪。针对HRM分析仪对温度控制和荧光检测的高分辨率要求，设计压控热管恒温槽构建高分辨热学系统控制新方法，基于光开关阵列高速切换的多路荧光检测技术实现高分辨快速荧光采集。通过微透镜、光纤耦合等制造工艺，使光学检测元件在结构上与热学模块无缝集成，同时保证温度控制和荧光检测的高分辨率，以保证仪器控温精度0.02℃、均匀性±0.05℃和多路荧光快速采集（100-200个数据/℃）的核心指标，研制出突变检测灵敏度达0.1%的首套自主知识产权的高性能HRM分析仪，并完成生物医学验证，为相关科学研究和原始创新提供新的技术手段。

中文关键词： 高分辨熔解曲线；荧光检测；温度控制；基因突变；单核苷酸多态性

英文摘要： High resolution melting curves analysis (HRM) is a new powerful tool for gene mutation and SNP detection. In order to solve the deficient of HRM analyzer in scientific research, we find the new mathematical empirical model, that using free-form algorithm to analyze the DNA melting process, and develop high throughput specialized HRM analyzer instrument. First by analyzing the detects of nearest neighbor thermodynamic model of double stand DNA melting, we employ the free-form algorithm to extract the feature of melting curves and find the complex physics laws of the process. Then new thermal system based on pressure controlled heat pipe and optical system based on high speed light switching array are developed to fulfill the requirements of high resolution temperature control and fluorescence detection. The thermal block and optical block are combined with micro-lens and optical fiber coupling engineer crafts. The key parameters of temperature and fluorescence for high resolution melting analysis are: the temperature control accuracy of 0.02℃, the temperature uniformity of ±0.05℃, and multi-channel fluorescence data density of 100-200 data/℃. Finally, a high-performance HRM analyzer with the first set of independent intellectual property rights is developed. The sensitivity of gene mutation and SNP detection will

英文关键词： High resolution melting；Fluorescence detection；Temperature control；Gene mutation；SNP