Integrated capture, concentration, polymerase chain reaction, and capillary electrophoretic analysis of pathogens on a chip.
Integrated capture, concentration, polymerase chain reaction, and capillary electrophoretic analysis of pathogens on a chip.
复制标题
DOI：
10.1021/ac900060r
复制
发表时间：
2009-05-01
影响因子：
7.4
通讯作者：
Mathies, Richard A.
中科院分区：
文献类型：
--
作者：
Beyor, Nathaniel;Yi, Lina;Seo, Tae Seok;Mathies, Richard A.
提出了一种用于病原体检测的芯片实验室系统，该系统集成了细胞预浓缩、纯化、PCR和毛细管电泳（CE）分析。该微型装置由微型泵和阀门、细胞捕获结构、100 nL PCR反应器和用于扩增子分离的5 cm长CE柱组成。引入10 - 100 μL的样品体积，并驱动其通过磁性约束免疫磁珠流化床，在流化床中捕获靶细胞。在细胞捕获后，使用芯片上泵将珠粒转移到PCR反应器中进行DNA扩增。PCR产物经毛细管电泳进样到毛细管电泳柱上进行分离和检测。coli K12和E. coliO 157靶基因。用E. coliO 157靶标，进样体积为50 μL。最后，建立了E. coliO 157在K12以1：1000的比例存在下的表达，说明了我们的系统在高浓度背景下识别靶细胞的能力。这种细胞捕获-PCR-CE微系统是开发用于病原体检测的快速、灵敏和特异性芯片实验室设备的重大进展。
A lab-on-a-chip system for pathogen detection is presented that integrates cell preconcentration, purification, PCR, and capillary electrophoretic (CE) analysis. The microdevice is comprised of micropumps and valves, a cell capture structure, a 100 nL PCR reactor, and a 5-cm long CE column for amplicon separation. Sample volumes ranging from 10 to 100 μL are introduced and driven through a fluidized bed of magnetically constrained immunomagnetic beads where the target cells are captured. After cell capture, beads are transferred using the on-chip pumps to the PCR reactor for DNA amplification. The resulting PCR products are electrophoretically injected onto the CE column for separation and detection of E. coli K12 and E. coli O157 targets. A detection limit of 0.2 cfu/μL is achieved using the E. coli O157 target and an input volume of 50 μL. Finally, the sensitive detection of E. coli O157 in the presence of K12 at a ratio of 1:1000 illustrates the capability of our system to identify target cells in a high commensal background. This cell capture-PCR-CE microsystem is a significant advance in the development of rapid, sensitive, and specific lab-on-a-chip devices for pathogen detection.
登录
查看更多内容
影响因子：
64.8
作者：
Perna, NT;Plunkett, G;Blattner, FR
通讯作者：
Blattner, FR
影响因子：
4.1
作者：
He, Y;Zhang, YHH;Yeung, ES
通讯作者：
Yeung, ES
影响因子：
7.4
作者：
SEILER, K;HARRISON, DJ;MANZ, A
通讯作者：
MANZ, A
影响因子：
4.2
作者：
Ocvirk, G;Tang, T;Harrison, DJ
通讯作者：
Harrison, DJ
影响因子：
7.4
作者：
Liu, Peng;Seo, Tae Seok;Mathies, Richard A.
通讯作者：
Mathies, Richard A.