Characterization of dynamic solid phase DNA extraction from blood with magnetically controlled silica beads

被引:40
作者
Duarte, Gabriela R. M. [1 ,4 ,5 ]
Price, Carol W. [1 ]
Littlewood, Janice L. [1 ]
Haverstick, Doris M. [2 ]
Ferrance, Jerome P. [1 ]
Carrilho, Emanuel [4 ,5 ]
Landers, James P. [1 ,2 ,3 ]
机构
[1] Univ Virginia, Dept Chem, Charlottesville, VA 22904 USA
[2] Univ Virginia, Hlth Sci Ctr, Dept Pathol, Charlottesville, VA 22908 USA
[3] Univ Virginia, Dept Mech Engn, Charlottesville, VA 22904 USA
[4] Univ Sao Paulo, Inst Quim Sao Carlos, BR-13566590 Sao Carlos, SP, Brazil
[5] Inst Nacl Ciencia & Tecnol Bioanalit, BR-13083970 Campinas, SP, Brazil
基金
巴西圣保罗研究基金会;
关键词
CHEMICAL-ANALYSIS SYSTEMS; NUCLEIC-ACIDS; PURIFICATION; MICROCHIP; MONOLITH; CAPTURE; AGAROSE; SAMPLES;
D O I
10.1039/b918996c
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A novel solid phase extraction technique is described where DNA is bound and eluted from magnetic silica beads in a manner where efficiency is dependent on the magnetic manipulation of the beads and not on the flow of solution through a packed bed. The utility of this technique in the isolation of reasonably pure, PCR-amplifiable DNA from complex samples is shown by isolating DNA from whole human blood, and subsequently amplifying a fragment of the beta-globin gene. By effectively controlling the movement of the solid phase in the presence of a static sample, the issues associated with reproducibly packing a solid phase in a microchannel and maintaining consistent flow rates are eliminated. The technique described here is rapid, simple, and efficient, allowing for recovery of more than 60% of DNA from 0.6 mu L of blood at a concentration which is suitable for PCR amplification. In addition, the technique presented here requires inexpensive, common laboratory equipment, making it easily adopted for both clinical point-of-care applications and on-site forensic sample analysis.
引用
收藏
页码:531 / 537
页数:7
相关论文
共 28 条
[21]  
WALKER JM, 1985, METHOD MOL BIOL, V2, DOI DOI 10.1385/0896030644
[22]   DNA extraction using a tetramethyl orthosilicate-grafted photopolymerized monolithic solid phase [J].
Wen, J ;
Guillo, C ;
Ferrance, JP ;
Landers, JP .
ANALYTICAL CHEMISTRY, 2006, 78 (05) :1673-1681
[23]   Purification of nucleic acids in microfluidic devices [J].
Wen, Jian ;
Legendre, Lindsay A. ;
Bienvenue, Joan M. ;
Landers, James P. .
ANALYTICAL CHEMISTRY, 2008, 80 (17) :6472-6479
[24]   Microfluidic chip-based protein capture from human whole blood using octadecyl (C18) silica beads for nucleic acid analysis from large volume samples [J].
Wen, Jian ;
Guillo, Christelle ;
Ferrance, Jerome P. ;
Landers, James P. .
JOURNAL OF CHROMATOGRAPHY A, 2007, 1171 (1-2) :29-36
[25]   Microfluidic-based DNA purification in a two-stage, dual-phase microchip containing a reversed-phase and a photopolymerized monolith [J].
Wen, Jian ;
Guillo, Christelle ;
Ferrance, Jerome P. ;
Landers, James P. .
ANALYTICAL CHEMISTRY, 2007, 79 (16) :6135-6142
[26]  
Wolfe KA, 2002, ELECTROPHORESIS, V23, P727, DOI 10.1002/1522-2683(200203)23:5<727::AID-ELPS727>3.0.CO
[27]  
2-O
[28]   Microchip-based macroporous silica sol-gel monolith for efficient isolation of DNA from clinical samples [J].
Wu, Qirong ;
Bienvenue, Joan M. ;
Hassan, Benjamin J. ;
Kwok, Yien C. ;
Giordano, Braden C. ;
Norris, Pamela M. ;
Landers, James P. ;
Ferrance, Jerome P. .
ANALYTICAL CHEMISTRY, 2006, 78 (16) :5704-5710