Integrated and finger-actuated microfluidic chip for point-of-care testing of multiple pathogens
Integrated and finger-actuated microfluidic chip for point-of-care testing of multiple pathogens
复制标题
用于多种病原体即时检测的集成手指驱动微流控芯片
DOI：
10.1016/j.talanta.2020.121844
复制
发表时间：
2021-03-01
期刊：
影响因子：
6.1
通讯作者：
Liu, Bi-Feng
中科院分区：
文献类型：
--
作者：
Chen, Peng;Chen, Chen;Liu, Bi-Feng
为了同时检测多种不同类型的细菌病原体，研制了基于凝胶的环介导等温扩增(GLAMP)和指动式微流控芯片(MU FAChip)的集成技术。开发的MU FA芯片由三个PDMS层组成，通过两个胶带连接在一起。上层PDMS层的多个腔室用于样品制备，底层PDMS层的相应腔室用于长期储存不含DNA模板的LAMP试剂。中间薄薄的PDMS层包含十字形的切割，就像手指驱动的流体控制阀门。为了减少芯片上的操作步骤，如移液和处理样本，Whatman CloneSaver卡预先嵌入到顶层腔室中，用于芯片上DNA的提取和纯化。只需简单地按下顶层，手指驱动的阀门就会打开，顶层的DNA样本就可以流入底层的反应室进行Glamp反应。对于POCT应用，分别在微型Peltier加热器和便携式荧光成像系统上进行了芯片上的灯反应和成像。在优化的条件下，可同时检测多种病原体，且具有较高的选择性和灵敏度(最低可达1.6个细胞)。开发的MU FAX芯片为基于GLAMP的病原体检测提供了一个快速且易于操作的平台，具有现场检测的潜力，特别是在资源有限的地区。
The integration of gel-based loop-mediated isothermal amplification (gLAMP) and finger-actuated microfluidic chip (mu FAchip) was developed for the simultaneous detection of various different types of bacterial pathogens. The developed mu FAchip consisted of three PDMS layers attached together by two adhesive tapes. Multiple chambers in the top PDMS layer were used for sample preparation, and the corresponding chambers in the bottom PDMS layer was used for long-term storage of LAMP reagents without DNA templates. The thin PDMS layer in the middle contained cross-shaped cuts as finger-actuated valves for fluid control. To reduce operation steps on the chip, such as pipetting and manipulation of samples, Whatman CloneSaver card was pre-embedded in the top chambers for on-chip DNA extraction and purification. Upon a simple press on the top layer, the finger-actuated valve was opened up, allowing DNA samples on the top layer flow into the bottom reaction chambers for gLAMP reaction. For POCT applications, on-chip LAMP reaction and imaging were conducted on a miniaturized peltier heater and a portable fluorescence imaging system respectively. Under the optimized condition, multiple pathogens were detected simultaneously with high selectivity and sensitivity (as low as 1.6 cells). The developed mu FAchip provided a rapid and easy-to-operate platform for gLAMP-based pathogen detection, with the potential for in-field detection, especially in areas with limited resources.