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使用表面改性硅柱阵列从全血中制备细菌DNA样本。

Bacterial DNA sample preparation from whole blood using surface-modified Si pillar arrays.

作者信息

Hwang Kyu-Youn, Lim Hee-Kyun, Jung Seong-Young, Namkoong Kak, Kim Joon-Ho, Huh Nam, Ko Christopher, Park Jae-Chan

机构信息

Bio & Health Lab., Samsung Advanced Institute of Technology (SAIT), Mt. 14-1, Nongseo-Dong, Giheung-Gu, Yongin-Si, Gyeonggi-Do, South Korea, 449-712.

出版信息

Anal Chem. 2008 Oct 15;80(20):7786-91. doi: 10.1021/ac8012048. Epub 2008 Aug 27.

DOI:10.1021/ac8012048
PMID:18729477
Abstract

A novel bacterial DNA sample preparation device for molecular diagnostics has been developed. On the basis of optimized conditions for bacterial adhesion, surface-modified silicon pillar arrays for bacterial cell capture were fabricated, and their ability to capture bacterial cells was demonstrated. The capture efficiency for bacterial cells such as Escherichia coli, Staphylococcus epidermidis, and Streptococcus mutans in buffer solution was over 75% with a flow rate of 400 microL/min. Moreover, the proposed method captured E. coli cells present in 50% whole blood effectively. The captured cells from whole blood were then in- situ lyzed on the surface of the microchip, and the eluted DNA was successfully amplified by qPCR. These results demonstrate that the full process of pathogen capture to DNA isolation from whole blood could be automated in a single microchip.

摘要

一种用于分子诊断的新型细菌DNA样本制备装置已被开发出来。基于优化的细菌粘附条件,制备了用于捕获细菌细胞的表面改性硅柱阵列,并展示了其捕获细菌细胞的能力。在缓冲溶液中,以400微升/分钟的流速,对大肠杆菌、表皮葡萄球菌和变形链球菌等细菌细胞的捕获效率超过75%。此外,所提出的方法能有效捕获50%全血中存在的大肠杆菌细胞。然后,将从全血中捕获的细胞在微芯片表面进行原位裂解,并通过qPCR成功扩增洗脱的DNA。这些结果表明,从全血中捕获病原体到分离DNA的整个过程可以在单个微芯片中实现自动化。

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