Real-time PCR detection of Listeria monocytogenes using an integrated microfluidics platform
Real-time PCR detection of Listeria monocytogenes using an integrated microfluidics platform
复制标题
DOI：
10.1016/j.snb.2004.10.022
复制
发表时间：
2005-05-27
影响因子：
8.4
通讯作者：
Batt, CA
中科院分区：
文献类型：
--
作者：
Cady, NC;Stelick, S;Batt, CA
一种小型化的、全自动的、基于PCR的检测系统已经被开发用于细菌病原体的快速检测。构建了单块DNA纯化/实时荧光PCR芯片，并对其纯化和检测单核细胞增生李斯特菌的能力进行了测试。使用二氧化硅包被的微结构，核酸可以被选择性地结合，洗涤和洗脱，用于随后的实时PCR。这些微结构被包含在一个集成的检测微芯片中，该芯片包含两个不同的区域，一个用于DNA纯化，一个用于实时PCR。利用集成微处理器、泵、阀、热循环仪和荧光检测模块的自动化检测平台，使用微芯片通过SYBR绿色荧光染料进行实时PCR扩增来纯化和检测细菌DNA。在10（4）和10（7）L之间。使用该系统可以检测单核细胞增多性细胞，平均周转时间为45分钟。（c）2004 Elsevier B. V.保留所有权利。
A miniaturized, fully automated, PCR-based detection system has been developed for the rapid detection of bacterial pathogens. Monolithic DNA purification/real-time PCR silicon chips were fabricated and tested for their ability to purify and detect the pathogeneic bacterium Listeria monocytogenes. Using silica-coated microstructures, nucleic acids could be selectively bound, washed and eluted for subsequent real-time PCR. These microstructures were included in an integrated detection microchip containing two distinct regions, one for DNA purification and one for real-time PCR. Using an automated detection platform with integrated microprocessor, pumps, valves, thermocycler and fluorescence detection modules, microchips were used to purify and detect bacterial DNA by real-time PCR amplification using SYBR Green fluorescent dye. Between 10(4) and 10(7) L. monocytogenes cells could be detected using this system with an average turnaround time of 45 min. (c) 2004 Elsevier B.V. All rights reserved.