Rapid detection of bacterial cell from whole blood: Integration of DNA sample preparation into single micro-PCR chip

被引:28
作者
Hwang, Kyu-Youn [1 ,2 ]
Jeong, Sung-Young [1 ]
Kim, Young-Rok [3 ,4 ]
Namkoong, Kak [1 ]
Lim, Hee-Kyun [1 ,5 ]
Chung, Won-Seok [1 ]
Kim, Joon-Ho [1 ]
Huh, Nam [1 ]
机构
[1] Samsung Elect Co, Bio & Hlth Lab, SAIT, Yongin 446712, Gyeonggi Do, South Korea
[2] Seoul Natl Univ, Interdisciplinary Program Bioengn, Seoul 151742, South Korea
[3] Kyung Hee Univ, Inst Life Sci & Resources, Yongin 446701, Gyeonggi Do, South Korea
[4] Kyung Hee Univ, Dept Food Sci & Biotechnol, Yongin 446701, Gyeonggi Do, South Korea
[5] Korea Adv Inst Sci & Technol, Dept Bio & Brain Engn, Coll Life Sci & Bioengn, Taejon 305701, South Korea
关键词
Micro-PCR; DNA sample preparation; Integration; Bacteria; POLYMERASE-CHAIN-REACTION; SILICON-GLASS CHIPS; MICROFLUIDIC DEVICES; PURIFICATION; AMPLIFICATION; SYSTEM;
D O I
10.1016/j.snb.2009.11.005
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A novel bacterial cell detection method from blood samples has been developed for molecular diagnostics. Functional integration of DNA sample preparation into polymerase chain reaction (PCR) chip enabled detection of pathogenic bacterial cells in a single microchip. Surface-modified micropillars possessing affinity for bacterial cells were fabricated inside a PCR chip, and reaction conditions were optimized to render the microchip with high surface-to-volume ratio PCR-compatible. After bacterial cells were captured on the micropillars from whole blood and PCR inhibitors were washed out. PCR mixture was injected to allow real-time amplification of DNA extracted from the isolated cells. Cell enrichment effect produced by volume reduction from large initial sample to small micro-PCR chip chamber led to increased detection sensitivity. Moreover, the developed method from sample preparation to detection of bacterial cells from whole blood took less than 1 h. These results demonstrated that the surface-modified pillar-packed microchip would be a practical approach for integration into Lab-On-a-Chip (LOC) to enable point-of-care genetic analysis. Crown Copyright (C) 2009 Published by Elsevier BM. All rights reserved.
引用
收藏
页码:46 / 51
页数:6
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