基于高通量微弱荧光快速检测的高分辨熔解曲线分析仪

高分辨熔解(HRM)曲线分析技术是近年发展起来的一种用于基因突变检测和单核苷酸多态性(SNP)分析的新方法,它通过实时监测PCR产物升温过程中双链DNA饱和染料的荧光强度变化来分析核酸序列的微小差异。根据HRM分析仪对荧光检测的时间和灵敏度需求,提出基于光开关阵列的多路高速荧光激发和检测模块实现高通量的微弱荧光快速检测;并根据HRM荧光数据特点,对原始荧光曲线进行滤波、基线探测、归一化和对温度微分等处理,从熔解曲线两端的线形区域自动提取基线作为归一化的标准,可以在不损失曲线形态特征信息的情况下获得更为精确的熔解温度,从而实现不同基因型熔解曲线的快速、准确识别。

High Resolution Melting Analyzer Based on High Throughout Fast Detection of Weak Fluorescence

High resolution melting curves analysis (HRM) is a recently developed powerful technique for the detection of gene mutations and SNP analysis. Monitoring the changes in fluorescence from saturated dye labeled on dsDNA, as a PCR products (amplicons) are melted, the minor difference of nucleic acid sequence can be discriminated. Here fast fluorescence detection system based on fiber-optics switch array is designed for fulfilling the requirements of speed and sensitivity. High-density fluorescence data are processed by a series of procedure, such as smoothed, automatic baseline detection, normalizing and differential to temperature, different gene types could be recognized quickly and correctly by the high resolution melting curves.