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
[1]   RAPID AND SIMPLE METHOD FOR PURIFICATION OF NUCLEIC-ACIDS [J].
BOOM, R ;
SOL, CJA ;
SALIMANS, MMM ;
JANSEN, CL ;
WERTHEIMVANDILLEN, PME ;
VANDERNOORDAA, J .
JOURNAL OF CLINICAL MICROBIOLOGY, 1990, 28 (03) :495-503
[2]   Nucleic acid purification using microfabricated silicon structures [J].
Cady, NC ;
Stelick, S ;
Batt, CA .
BIOSENSORS & BIOELECTRONICS, 2003, 19 (01) :59-66
[3]   Chitosan as a polymer for pH-induced DNA capture in a totally aqueous system [J].
Cao, Weidong ;
Easley, Christopher J. ;
Ferrance, Jerome P. ;
Landers, James P. .
ANALYTICAL CHEMISTRY, 2006, 78 (20) :7222-7228
[4]   AN INEXPENSIVE AND SIMPLE METHOD FOR DNA PURIFICATIONS ON SILICA PARTICLES [J].
CARTER, MJ ;
MILTON, ID .
NUCLEIC ACIDS RESEARCH, 1993, 21 (04) :1044-1044
[5]   RECOVERY OF DNA SEGMENTS FROM AGAROSE GELS [J].
CHEN, CW ;
THOMAS, CA .
ANALYTICAL BIOCHEMISTRY, 1980, 101 (02) :339-341
[6]   Rapid, automated nucleic acid probe assays using silicon microstructures for nucleic acid concentration [J].
Christel, LA ;
Petersen, K ;
McMillan, W ;
Northrup, MA .
JOURNAL OF BIOMECHANICAL ENGINEERING-TRANSACTIONS OF THE ASME, 1999, 121 (01) :22-27
[7]   Developments toward a complete micro-total analysis system for Duchenne muscular dystrophy diagnosis [J].
Ferrance, JP ;
Wu, QR ;
Giordano, B ;
Hernandez, C ;
Kwok, Y ;
Snow, K ;
Thibodeau, S ;
Landers, JP .
ANALYTICA CHIMICA ACTA, 2003, 500 (1-2) :223-236
[8]  
HENGEN P, 1995, MOL BIOL CURRENT INN
[9]  
Legendre L.A., 2008, JALA - Journal of the Association for Laboratory Automation, V13, P351, DOI DOI 10.1016/J.JALA.2008.08.001
[10]  
LEGENDRE LA, 2009, UNPUB