Sample preparation module for bacterial lysis and isolation of DNA from human urine

Biomed Microdevices. 2009 Jun;11(3):671-678. doi: 10.1007/s10544-008-9277-1.

Abstract

Silica impregnated polymer monolithic columns may provide a simple method for lysing and extracting DNA from bacteria inside of microfluidic chips. Here we use Escherichia coli as a test organism for a point of care thermoplastic microfluidic module designed to take in a urine sample, mix it with lysis buffer, and perform a hybrid chemical/mechanical lysis and solid phase extraction of nucleic acids from the sample. To demonstrate proof-of-concept, we doped human hematuric urine samples with E. coli at concentrations ranging from 10(1)-10(5) colony-forming units/mL (CFU/mL) to simulate patient samples. We then performed on-chip lysis and DNA extraction. The bacterial DNA was amplified using real-time PCR demonstrating lysis and isolation down to 10(1) CFU/mL. Results were comparable to a commercial kit at higher concentrations and performed better at recovering DNA at lower concentrations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteriolysis*
  • Biological Assay
  • Computer Simulation
  • DNA, Bacterial / isolation & purification*
  • Disposable Equipment
  • Escherichia coli / genetics
  • Green Fluorescent Proteins / genetics
  • Humans
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods*
  • Microscopy, Electron, Scanning
  • Miniaturization
  • Plasmids
  • Polymerase Chain Reaction
  • Reagent Kits, Diagnostic
  • Silicon Dioxide / chemistry
  • Urine / microbiology*

Substances

  • DNA, Bacterial
  • Reagent Kits, Diagnostic
  • Green Fluorescent Proteins
  • Silicon Dioxide