One-step pathogen specific DNA extraction from whole blood on a centrifugal microfluidic device.
We report a fully integrated, pathogen-specific DNA extraction device utilizing centrifugal microfluidics on a polymer based CD platform. By use of the innovative laser irradiated Ferrowax microvalve (LIFM) together with the rapid cell lysis method using laser irradiation on magnetic particles, we could, for the first time, demonstrate a fully integrated pathogen specific DNA extraction from whole blood on a CD. As a model study, DNA extraction experiments from whole blood spiked with Hepatitis B virus (HBV) and E.coli were conducted. The total process of the plasma separation, mixing with magnetic beads conjugated with target specific antibodies, removal of plasma residual, washing and DNA extraction was finished within 12 min with only one manual step, the loading of 100 microL of whole blood. Real-time PCR results showed that the concentration of DNA prepared on a CD using a portable sample preparation device was as good as those by conventional bench top protocol. It demonstrates that our novel centrifugal microfluidics platform enables a full integration of complex biological reactions that require multi-step fluidic control.
Cho
Yoon-Kyoung
YK
Bio Device Research Laboratory, Samsung Advanced Institute of Technology, P.O. Box 111, Suwon, 440-600, Korea.
Lee
Jeong-Gun
JG
Park
Jong-Myeon
JM
Lee
Beom-Seok
BS
Lee
Youngsun
Y
Ko
Christopher
C
D002498:Centrifugation / Q000295:instrumentation* / Q000379:methods*; D004269:DNA, Bacterial / Q000097:blood* / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000472:pathogenicity; D006801:Humans; D008280:Magnetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*
D004269:DNA, Bacterial