Validation of a centrifugal microfluidic sample lysis and homogenization platform for nucleic acid extraction with clinical samples.
The applications of microfluidic technologies in medical diagnostics continue to increase, particularly in the field of nucleic acid diagnostics. While much attention has been focused on the development of nucleic acid amplification and detection platforms, sample preparation is often taken for granted or ignored all together. Specifically, little or no consideration is paid to the development of microfluidic systems that efficiently extract nucleic acids from biological samples. Here, a centrifugal microfluidic platform for mechanical sample lysis and homogenization is presented. The system performs sample lysis through a magnetically actuated bead-beating system followed by a centrifugal clarification step. The supernatant is then transferred for extraction using a unique siphon. Several other new microfluidic functions are implemented on this centrifugal platform as well, including sample distribution, a unique hydraulic capillary valve, and self-venting. Additionally, the improved system has features with a small footprint designed specifically for integration with further downstream processing steps. Biological validation of the platform is performed using Bacillus subtilis spores and clinical samples (nasopharyngeal aspirates) for respiratory virus detection. The platform was found to be as efficient as in-tube bead-beating lysis and homogenization for nucleic acid extraction, and capable of processing 4 samples in batch to near PCR-ready products in under 6 min.
Siegrist
Jonathan
J
University of California, Irvine, Irvine, CA 92697, USA. jsiegris@uci.edu
Gorkin
Robert
R
Bastien
Martine
M
Stewart
Gale
G
Peytavi
Régis
R
Kido
Horacio
H
Bergeron
Michel
M
Madou
Marc
M
D002458:Cell Fractionation / Q000295:instrumentation*; D002498:Centrifugation / Q000295:instrumentation*; D005591:Chemical Fractionation; D017076:Computer-Aided Design; D004867:Equipment Design; D019544:Equipment Failure Analysis; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D009696:Nucleic Acids / Q000302:isolation & purification*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000295:instrumentation*
D009696:Nucleic Acids