An integrated disposable device for DNA extraction   and helicase dependent amplification.
Here we report the demonstration of an integrated microfluidic chip that performs helicase dependent amplification (HDA) on samples containing live bacteria. Combined chip-based sample preparation and isothermal amplification are attractive for world health applications, since the need for instrumentation to control flow rate and temperature changes are reduced or eliminated. Bacteria lysis, nucleic acid extraction, and DNA amplification with a fluorescent reporter are incorporated into a disposable polymer cartridge format. Smart passive fluidic control using a flap valve and a hydrophobic vent (with a nanoporous PTFE membrane) with a simple on-chip mixer eliminates multiple user operations. The device is able to detect as few as ten colony forming units (CFU) of E. coli in growth medium.
Mahalanabis
Madhumita
M
Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.
Do
Jaephil
J
ALMuayad
Hussam
H
Zhang
Jane Y
JY
Klapperich
Catherine M
CM
D001419:Bacteria / Q000235:genetics / Q000378:metabolism; D003470:Culture Media; D004247:DNA / Q000235:genetics*; D004265:DNA Helicases / Q000235:genetics* / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D005243:Feces; D044085:Microfluidics / Q000295:instrumentation*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation; D013696:Temperature
D003470:Culture Media; D004247:DNA; C070331:Tli polymerase; D004259:DNA-Directed DNA Polymerase; D004265:DNA Helicases