Validation of a blood plasma separation system by biomarker detection.
A microfluidic system was developed for blood plasma separation at high flow rate. This system uses only hydrodynamic forces to separate plasma from whole blood. The microfluidic network features a series of constrictions and bifurcations to enhance the product yield and purity. A maximum purity efficiency of 100% is obtained on blood with entrance hematocrit level up to 30% with a flow rate of 2 mL h(-1). Flow cytometry was performed on the extracted plasma to evaluate the separation efficiency and to assess cell damage. A core target of this study was the detection of cell-free DNA from the on-chip extracted plasma. To this effect, PCR was successfully carried out off-chip on the cell-free DNA present in the plasma extracted on-chip. A house-keeping gene sequence (GAPDH) was amplified without the need for a purification after the separation, thereby showing the high quality of the plasma sample. The resulting data suggests that the system can be used as a preliminary module of a total analysis system for cell-free DNA detection in human plasma.
Kersaudy-Kerhoas
Maïwenn
M
MIcroSystems Engineering Centre (MISEC), School of Engineering & Physical Sciences, Heriot-Watt University, Earl Mountbatten Building, Edinburgh, United Kingdom. mk87@hw.ac.uk
Kavanagh
Deirdre M
DM
Dhariwal
Resham S
RS
Campbell
Colin J
CJ
Desmulliez
Marc P Y
MP
D015415:Biomarkers / Q000097:blood; D005591:Chemical Fractionation / Q000295:instrumentation / Q000379:methods*; D004247:DNA / Q000097:blood / Q000235:genetics; D004586:Electrophoresis; D005434:Flow Cytometry; D006400:Hematocrit; D006801:Humans; D007700:Kinetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D010949:Plasma; D016133:Polymerase Chain Reaction
D015415:Biomarkers; D004247:DNA