基于微流控荧光定量PCR的MRSA快速检测技术研究

摘要

针对耐甲氧西林金黄色葡萄球菌（MRSA）检测的问题，发展一种基于微流控荧光定量PCR的致病菌快速检测新技术。采用光刻法制作了微流控PCR芯片，研制了集成高性能温度控制模块和高灵敏度荧光检测模块的微流控荧光定量PCR分析系统。通过检测特异性基因femA和mecA对MRSA进行快速鉴别。实验结果表明，使用微流控PCR芯片可以成功实现MRSA特异性基因的快速检测，相对传统的管式PCR，芯片使用6µL试剂在56min完成了MRSA特异基因的检测，不仅节约了反应试剂，而且极大提高了检测速度。该技术可扩展到其他致病菌的检测，具有良好的应用前景。%For Methicillin-resistant Staphylococcus Aureus (MRSA) identification, a new rapid detection technology of pathogenic bacteria was developed using microfluidic real-time PCR. The detection was conducted on a microlfuidic chip fabricated through soft lithography method. A microlfuidic real-time PCR analysis system,which integrated a high-performance temperature control module and a high-sensitivity lfuorescence detection unit,was developed. Speciifc genes femA and mecA are ampliifed on a microlfuidic chip for fast identiifcation of MRSA. The experimental results showed that the microlfuidic PCR chip could achieve rapid detection of speciifc MRSA genes in 56 min with 6 µL reagent. Comparing with the traditional tube PCR, this microlfuidic PCR technology is reagent saving and greatly improving the speed of the detection. The system presented here can be extended to other pathogens’ detection and will lead to a promising application prospect.