23 January 2006 Rapid and automated sample preparation for nucleic acid extraction on a microfluidic CD (compact disk)
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Abstract
Rapid and automated preparation of PCR (polymerase chain reaction)-ready genomic DNA was demonstrated on a multiplexed CD (compact disk) platform by using hard-to-lyse bacterial spores. Cell disruption is carried out while beadcell suspensions are pushed back and forth in center-tapered lysing chambers by angular oscillation of the disk - keystone effect. During this lysis period, the cell suspensions are securely held within the lysing chambers by heatactivated wax valves. Upon application of a remote heat to the disk in motion, the wax valves release lysate solutions into centrifuge chambers where cell debris are separated by an elevated rotation of the disk. Only debris-free DNA extract is then transferred to collection chambers by capillary-assisted siphon and collected for heating that inactivates PCR inhibitors. Lysing capacity was evaluated using a real-time PCR assay to monitor the efficiency of Bacillus globigii spore lysis. PCR analysis showed that 5 minutes' CD lysis run gave spore lysis efficiency similar to that obtained with a popular commercial DNA extraction kit (i.e., IDI-lysis kit from GeneOhm Sciences Inc.) which is highly efficient for microbial cell and spore lysis. This work will contribute to the development of an integrated CD-based assay for rapid diagnosis of infectious diseases.
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Jitae Kim, Horacio Kido, Jim V. Zoval, Dominic Gagné, Régis Peytavi, François J. Picard, Martine Bastien, Maurice Boissinot, Michel G. Bergeron, Marc J. Madou, "Rapid and automated sample preparation for nucleic acid extraction on a microfluidic CD (compact disk)", Proc. SPIE 6112, Microfluidics, BioMEMS, and Medical Microsystems IV, 611204 (23 January 2006); doi: 10.1117/12.646932; https://doi.org/10.1117/12.646932
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