Sample introduction interface for on-chip nucleic acid-based analysis of Helicobacter pylori from stool samples
Sample introduction interface for on-chip nucleic acid-based analysis of Helicobacter pylori from stool samples
复制标题
DOI：
10.1039/c6lc00228e
复制
发表时间：
2016-01-01
期刊：
影响因子：
6.1
通讯作者：
Shaw, K. J.
中科院分区：
文献类型：
--
作者：
Mosley, O.;Melling, L.;Shaw, K. J.
尽管近年来基于微流体的集成诊断系统取得了进展，但样品导入界面，特别是涉及大体积样品时，经常被忽视。我们提出了一个样品导入接口，允许直接在芯片上处理粗粪便样品，以检测幽门螺杆菌（H. pylori）。采用了IFAST（表面张力辅助的非混相过滤）的原理，包括一个大容量的样品室，带有一个基于隔膜的界面，用于粪便样品导入。固体朝向盐和干燥的超顺磁颗粒（pmp）可以存储在芯片上，并在样品添加后重建，从而简化了从幽门螺杆菌细胞释放DNA及其与pmp结合的过程。最后，pmp通过磁铁通过含有不混溶油溶液的洗涤室，进入洗脱室，在那里DNA被释放到水介质中进行后续分析。整个过程只需要7分钟，同时使原油生物样品的工作体积减少了40倍。真实世界界面和快速DNA提取的结合为该方法在护理点（POC）设备中使用提供了潜力。
Despite recent advances in microfluidic-based integrated diagnostic systems, the sample introduction interface, especially with regards to large volume samples, has often been neglected. We present a sample introduction interface that allows direct on-chip processing of crude stool samples for the detection of Helicobacter pylori (H. pylori). The principle of IFAST (immiscible filtration assisted by surface tension) was adapted to include a large volume sample chamber with a septum-based interface for stool sample introduction. Solid chaotropic salt and dry superparamagnetic particles (PMPs) could be stored on-chip and reconstituted upon sample addition, simplifying the process of release of DNA from H. pylori cells and its binding to the PMPs. Finally, the PMPs were pulled via a magnet through a washing chamber containing an immiscible oil solution and into an elution chamber where the DNA was released into aqueous media for subsequent analysis. The entire process required only 7 min while enabling a 40-fold reduction in working volume from crude biological samples. The combination of a real-world interface and rapid DNA extraction offers the potential for the methodology to be used in point-of-care (POC) devices.