Increasing PCR sensitivity by removal of polymerase inhibitors in environmental samples by using dielectrophoresis

被引：20

作者：

del Carmen Jaramillo, Maria ^{[1
]}

Martinez-Duarte, Rodrigo ^{[2
]}

Huettener, Mario ^{[1
]}

Renaud, Philippe ^{[2
]}

Torrents, Eduard ^{[1
]}

Juarez, Antonio ^{[1
,3
]}

机构：

[1] Inst Bioengn Catalonia IBEC, Barcelona 08028, Spain

[2] Ecole Polytech Fed Lausanne, Microsyst Lab LMIS4, CH-1015 Lausanne, Switzerland

[3] Univ Barcelona, Dept Microbiol, Fac Biol, E-08028 Barcelona, Spain

来源：

BIOSENSORS & BIOELECTRONICS | 2013年 / 43卷

关键词：

Yeast; Dielectrophoresis; PCR; Separation; Carbon-electrode; CONTINUOUS SEPARATION; CELLS; MICROORGANISMS; PARTICLES; IMPEDANCE; MIXTURES;

D O I：

10.1016/j.bios.2012.12.049

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

Dielectrophoresis (DEP) is a powerful tool to manipulate cells and molecules in microfluidic chips. However, few practical applications using DEP exist. An immediate practical application of a carbon-electrode DEP system, in removing PCR inhibitors from a sample, is reported in this work. We use a high throughput carbon-electrode DEP system to trap yeast cells from a natural sample (fermented grape must) and then in situ remove contaminants that interfere with PCR analysis. Retrieval of this enriched and purified yeast population from the DEP system then allows for a significant increase of sensitivity during PCR analysis. Furthermore, the fact that DEP can discriminate between viable and non-viable cells minimizes the number of false positives commonly obtained when using PCR alone. Experimental results provide clear evidence that the carbon-electrode DEP-based sample preparation step can readily and effectively clean environmental samples from natural contaminants and improve PCR sensitivity. (c) 2012 Elsevier B.V. All rights reserved.

引用

页码：297 / 303

页数：7