痕量样品高灵敏度快速测量方法与便携式系统研究

为了实现高灵敏度、快速、准确鉴定痕量病原菌核酸样品,介绍了一种痕量核酸样品高灵敏度快速测量方法,并发展了一种微流控芯片核酸等温扩增实时分子诊断技术,研制了新型微纳体系生化反应载体芯片。通过表面惰性处理降低芯片表面对生物分子的吸附影响,构建一种大数值孔径、长工作距离的便携式共焦光学检测系统,有效消除背景荧光的影响,提高了检测灵敏度。在微纳升(7μL～40nL)试剂消耗反应体系水平,实现检测灵敏度5个DNA分子拷贝数,并以呼吸道感染疾病的大肠埃希氏菌检测为例,开展临床应用研究,满足低成本临床医疗应用需要。

Study of High Sensitive and Fast Detection to Trace Sample and Portable System

In order to carry out the goal of highly sensitive, fast and accurate detection and identification of pathogen, a novel method for highly sensitive and fast detecting trace amount of nucleic acid sample is introduced. A microfluidic chip isothermal DNA amplification for bacteria molecular diagnostics is developed. New micro-nanoliter reaction assay chip is manufactured, and the surface inertia processing technique of chip is advanced to drop the chip surface adsorption to DNA molecules. A portable confocal detector with high numerical aperture and long working distance is built to reduce background fluorescence and improve the sensitivity. The sensitivity of 5 DNA copies in micro-nanoliter reaction assay is obtained. The method and detector was verified by Escherichia coli identification to acute respiratory disease, and can meet the application of low-cost detection of clinic medicine.