New detection modality for label-free quantification of DNA in biological samples via superparamagnetic bead aggregation.

Combining DNA and superparamagnetic beads in a rotating magnetic field produces multiparticle aggregates that are visually striking, enabling label-free optical detection and quantification of DNA at levels in the picogram per microliter range. DNA in biological samples can be quantified directly by simple analysis of optical images of microfluidic wells placed on a magnetic stirrer without prior DNA purification. Aggregation results from DNA/bead interactions driven either by the presence of a chaotrope (a nonspecific trigger for aggregation) or by hybridization with oligonucleotides on functionalized beads (sequence-specific). This paper demonstrates quantification of DNA with sensitivity comparable to that of the best currently available fluorometric assays. The robustness and sensitivity of the method enable a wide range of applications, illustrated here by counting eukaryotic cells. Using widely available and inexpensive benchtop hardware, the approach provides a highly accessible low-tech microscale alternative to more expensive DNA detection and cell counting techniques.

[1]  Daniel C Leslie,et al.  Nucleic acid extraction techniques and application to the microchip. , 2009, Lab on a chip.

[2]  M. Dijkstra,et al.  Phase diagram of colloidal spheres in a biaxial electric or magnetic field. , 2010, The Journal of chemical physics.

[3]  T. Lion,et al.  Detection of gene expression by PCR amplification of RNA derived from frozen heparinized whole blood. , 1991, Nucleic acids research.

[4]  Chee Yoon Yue,et al.  Thermal bonding of PMMA: effect of polymer molecular weight , 2009 .

[5]  S. Biswal,et al.  Probing the stability of magnetically assembled DNA-linked colloidal chains. , 2009, Langmuir : the ACS journal of surfaces and colloids.

[6]  Marc Fermigier,et al.  Permanently Linked Monodisperse Paramagnetic Chains , 1998 .

[7]  Paul E. Otto MagneSil™ Paramagnetic Particles: Magnetics for DNA Purification , 2002 .

[8]  M. Burns,et al.  Modelling the limit of detection in real-time quantitative PCR , 2008 .

[9]  W. Al-Soud,et al.  Identification and Characterization of Immunoglobulin G in Blood as a Major Inhibitor of Diagnostic PCR , 2000, Journal of Clinical Microbiology.

[10]  Martin A. M. Gijs,et al.  Magnetic bead handling on-chip: new opportunities for analytical applications , 2004 .

[11]  Viswanadham Garimella,et al.  Homogeneous detection of unamplified genomic DNA sequences based on colorimetric scatter of gold nanoparticle probes , 2004, Nature Biotechnology.

[12]  L. Blum,et al.  DNA biosensors and microarrays. , 2008, Chemical reviews.

[13]  Daniel C Leslie,et al.  Size‐based separations as an important discriminator in development of proximity ligation assays for protein or organism detection , 2010, Electrophoresis.

[14]  R. Turner,et al.  Driving Forces for DNA Adsorption to Silica in Perchlorate Solutions , 1996 .

[15]  C. Mirkin,et al.  Scanometric DNA array detection with nanoparticle probes. , 2000, Science.

[16]  A. Akane,et al.  Identification of the heme compound copurified with deoxyribonucleic acid (DNA) from bloodstains, a major inhibitor of polymerase chain reaction (PCR) amplification. , 1994, Journal of forensic sciences.

[17]  J. Baudry,et al.  Flexible magnetic filaments as micromechanical sensors. , 2003, Physical review letters.

[18]  D Frenkel,et al.  Field-induced self-assembly of suspended colloidal membranes. , 2009, Physical review letters.

[19]  J. Storhoff,et al.  Selective colorimetric detection of polynucleotides based on the distance-dependent optical properties of gold nanoparticles. , 1997, Science.

[20]  Vishnu Swarup,et al.  Circulating (cell‐free) nucleic acids – A promising, non‐invasive tool for early detection of several human diseases , 2007, FEBS letters.

[21]  J. Bibette,et al.  Self-assembled magnetic nanowires made irreversible by polymer bridging. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[22]  Edward J. Wood,et al.  Biochemistry (3rd ed.) , 2004 .

[23]  K. A. Wolfe,et al.  Microchip-based purification of DNA from biological samples. , 2003, Analytical chemistry.

[24]  John M. Butler,et al.  Forensic DNA Typing: Biology, Technology, and Genetics of STR Markers , 2001 .

[25]  G. Kershaw,et al.  A Beginner’s Guide to Blood Cells , 1997 .

[26]  C. Mirkin,et al.  Homogeneous, Nanoparticle-Based Quantitative Colorimetric Detection of Oligonucleotides , 2000 .

[27]  J. Baudry,et al.  Magnetic force probe for nanoscale biomolecules. , 2005, Physical review letters.

[28]  Yasuhiro Sakamoto,et al.  Magnetic field-induced assembly of oriented superlattices from maghemite nanocubes , 2007, Proceedings of the National Academy of Sciences.

[29]  A. Gast,et al.  Structure evolution in a paramagnetic latex suspension , 1992 .

[30]  A. Gast,et al.  Mechanics of semiflexible chains formed by poly(ethylene glycol)-linked paramagnetic particles. , 2003, Physical review. E, Statistical, nonlinear, and soft matter physics.

[31]  Huixiang Li,et al.  Colorimetric detection of DNA sequences based on electrostatic interactions with unmodified gold nanoparticles. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Drew Provan A Beginner's Guide to Blood Cells, 2nd edn , 2004 .