目前临床病原菌检测主要依赖细菌培养的方法，该方法周期长、准确性低，且容易产生交叉污染和人体感染等问题。提出微流控芯片核酸恒温扩增分子诊断技术，开发离心进样空气隔离微流控芯片和共焦面成像光学检测系统，采用旋转扫描信号采集方法和薄层空气浴流动加热比例-积分-微分温控方法对呼吸道病原菌进行实验和分析。研制恒温扩增微流控芯片核酸分析系统，将系统检测灵敏度提高至10 copies，降低样品试剂消耗量至0.94 μL，在45 min内能够同时进行多种病原菌指标的并行分析鉴定。通过100例临床样品实验测试，得到所提系统与传统聚合酶链反应所得结果的总符合率大于98%，可满足医院、社区医疗、乡镇卫生诊所等低成本精准医疗应用的需要。

Molecular diagnostics is one of the most important tools currently in use for clinical pathogen detection due to its high sensitivity, specificity, and low consume of sample and reagent is keyword to low cost molecular diagnostics. In this paper, a sensitive DNA isothermal amplifi-cation method for fast clinical infectious diseases diagnostics at aM concentrations of DNA was developed using a polycarbonate (PC) microfluidic chip. A portable confocal optical fluo-rescence detector was specifically developed for the microfluidic chip that was capable of highly sensitive real-time detection of amplified products for sequence-specific molecular identification near the optical diffraction limit with low background. The molecular diagnostics of Listeria monocytogenes with nucleic acid extracted from stool samples was performed at a minimum DNA template concentration of 3.65 aM, and a detection limit of less than five copies of genomic DNA. Contrast to the general polymerase chain reaction (PCR) at eppendorf (EP) tube, the detection time in our developed method was reduced from 1.5 h to 45 min for multi-target parallel detection, the consume of sample and reagent was dropped from 25 L to 1.45 L. This novel microfluidic chip system and method can be used to develop a micro total analysis system as a clinically relevant pathogen molecular diagnostics method via the amplification of targets, with potential applications in biotechnology, medicine, and clinical molecular diagnostics.

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。