title: An integrated disposable device for DNA extraction   and helicase dependent amplification.
abstract: 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.
authors.affiliation: Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.
authors.forename: Madhumita
authors.initials: M
authors.lastname: Mahalanabis
authors.forename: Jaephil
authors.initials: J
authors.lastname: Do
authors.forename: Hussam
authors.initials: H
authors.lastname: ALMuayad
authors.forename: Jane Y
authors.initials: JY
authors.lastname: Zhang
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Klapperich
mesh_terms: 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
chemical_list: D003470:Culture Media; D004247:DNA; C070331:Tli polymerase; D004259:DNA-Directed DNA Polymerase; D004265:DNA Helicases