Paper-based
microfluidic analytical devices (μPADs) have
recently attracted attention as a point-of-care test kit because of
their low cost and nonrequirement for external forces. To directly
detect biomarkers in whole blood, however, they need to be assembled
with a filter such as a plasma separation membrane (PSM) because the
color of the blood cells interferes with the colorimetric assay. However,
this assembly process is rather complicated and cumbersome, and the
fluid does not uniformly move to the detection zone when the adhesion
between the paper and PSM is not perfect. In this study, we report
a simple three-dimensional (3D) printing method for fabricating PSM-integrated
3D-μPADs made of plastics without the need for additional assembly.
In detail, PSM was coated with parylene C to prevent its dissolution
from organic solvent during 3D printing. Then, the coated PSM was
superimposed on the paper. Detection zones and a reservoir were printed
on the paper and PSM via liquid photopolymerization, using a digital
light processing printer. The limit of detection of the PSM-integrated
3D-μPADs for glucose in whole blood was 0.3 mM, and these devices
demonstrated clinically relevant performance on diabetes patient blood
samples. Our 3D-μPADs can also simultaneously detect multiple
metabolic disease markers including glucose, cholesterol, and triglycerides
in whole blood. Our results suggest that our printing method is useful
for fabricating 3D-μPADs integrated with PSM for the direct
detection of biomarkers in whole blood.