Integrated microfluidic systems for sample preparation and detection of respiratory pathogen Bordetella pertussis.
An integrated microfluidic system for combined manipulation, pre-concentration, and lysis of samples containing Bordetella pertussis by dielectrophoresis and electroporation has been developed and implemented. The microfluidic device was able to pre-concentrate the amount of B. pertussis cells present in 200 microl of a B. pertussis suspension stock into a 20 microl volume. The device exhibited optimal sample pre-concentration of 6.7x at a stock value of 10(3) cfu/ml and at a flow rate of 250 microl/h. Electro-disruption experiments showed that on-chip-based electroporation is an effective solution for lysis of B. pertussis cells that is easily integrated with dielectrophoresis assisted pre-concentration procedures. Pulsed voltage applied, number of pulses, and presence of potassium chloride in a B. pertussis suspension showed a reduction in B. pertussis cell viability by electroporation; and transmission electron microscopy confirmed B. pertussis cell disruption by electroporation. Genetic amplification and detection of the pre-concentrated sample employing an integrated chip-based system demonstrated a complete chip approach for pathogen detection.
de la Rosa
Carlos
C
Schulich School of Engineering, Department of Electrical and Computer Engineering, 2500 University Drive, NW. Calgary, Alberta, Canada. T2N 1N4.
Prakash
Ranjit
R
Tilley
Peter A
PA
Fox
Julie D
JD
Kaler
Karan V i S
KV
D001886:Bordetella pertussis / Q000302:isolation & purification*; D015169:Colony Count, Microbial / Q000295:instrumentation* / Q000379:methods; D004586:Electrophoresis / Q000295:instrumentation* / Q000379:methods; D018274:Electroporation / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000295:instrumentation* / Q000379:methods; D018511:Systems Integration; D014917:Whooping Cough / Q000175:diagnosis* / Q000382:microbiology*