La. Christel et al., Rapid, automated nucleic acid probe assays using silicon microstructures for nucleic acid concentration, J BIOMECH E, 121(1), 1999, pp. 22-27
Citations number
13
Categorie Soggetti
Multidisciplinary
Journal title
JOURNAL OF BIOMECHANICAL ENGINEERING-TRANSACTIONS OF THE ASME
A system for rapid point-of-use nucleic acid (NA) analysis based on PCR tec
hniques is described. The extraction and concentration of DNA from test sam
ples has been accomplished utilizing silicon fluidic microchips with high s
urface-area-to-volume ratios. Short (500 bp) and medium size (48,000 bp) DN
A have been captured, washed, and eluted using the silicon dioxide surfaces
of these chips. Chaotropic (GuHCl) salt solutions were used as binding age
nts. Wash and elution agents consisted of ethanol-based solutions and water
, respectively. DNA quantities approaching 40 ng/cm(2) of binding area were
captured from input solutions in the 100-1000 ng/mL concentration range. F
or dilute samples of interest for pathogen detection, PCR and gel electroph
oresis were used to demonstrate extraction efficiencies of about 50 percent
, and concentration factors of about 10x using bacteriophage lambda DNA as
the target Rapid, multichannel PCR thermal cycling modules with integrated
solid-state detection components have also been demonstrated. These results
confirm the viability of utilizing these components as elements of a compa
ct, disposable cartridge system for the detection of NA in applications suc
h as clinical diagnostics, biowarfare agent detection, food quality control
, and environmental monitoring.