光学学报, 2012, 32 (2): 0217003, 网络出版: 2012-01-12   

痕量样品高灵敏度快速测量方法与便携式系统研究

Study of High Sensitive and Fast Detection to Trace Sample and Portable System
作者单位
1 清华大学医学院生物医学工程系, 北京 100084
2 生物芯片北京国家工程研究中心, 北京 102206
摘要
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(7 μL~40 nL)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。
Abstract
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.
参考文献

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黄国亮, 田浩, 李志永, 王同舟, 罗贤波, 马丽, 王璨, 邓涛. 痕量样品高灵敏度快速测量方法与便携式系统研究[J]. 光学学报, 2012, 32(2): 0217003. Huang Guoliang, Tian Hao, Li Zhiyong, Wang Tongzhou, Luo Xianbo, Ma Li, Wang Can, Deng Tao. Study of High Sensitive and Fast Detection to Trace Sample and Portable System[J]. Acta Optica Sinica, 2012, 32(2): 0217003.

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