Integrated microfluidic systems for sample preparation and detection of respiratory pathogen Bordetella pertussis
Carlos de la Rosa1, Ranjit Prakash, Peter A Tilley
1Schulich School of Engineering, Department of Electrical and Computer Engineering, 2500 University Drive, NW. Calgary, Alberta, Canada. T2N 1N4.
Abstract:
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.
Insights
This study presents an integrated microfluidic system for Bordetella pertussis detection. The device efficiently pre-concentrates and lyses bacteria using dielectrophoresis and electroporation for pathogen analysis.
Area of Science:
- Biotechnology
- Microfluidics
- Pathogen Detection
Background:
- Bordetella pertussis detection requires efficient sample preparation.
- Current methods for sample manipulation and lysis can be time-consuming and complex.
Purpose of the Study:
- To develop an integrated microfluidic system for combined manipulation, pre-concentration, and lysis of Bordetella pertussis.
- To demonstrate a complete chip-based approach for pathogen detection.
Main Methods:
- Development of a microfluidic device utilizing dielectrophoresis for pre-concentration and electroporation for cell lysis.
- Optimization of pre-concentration parameters including flow rate and initial cell concentration.
- Evaluation of electroporation effectiveness on Bordetella pertussis viability and cell disruption.
Main Results:
- The microfluidic device achieved a 6.7x pre-concentration of Bordetella pertussis cells from 200 microl to 20 microl at optimal conditions.
- On-chip electroporation effectively lysed Bordetella pertussis cells, confirmed by transmission electron microscopy.
- Integrated system demonstrated successful genetic amplification and detection of pre-concentrated pathogens.
Conclusions:
- The integrated microfluidic system offers an efficient and complete solution for Bordetella pertussis sample preparation and detection.
- Dielectrophoresis and electroporation are effective tools for on-chip manipulation and lysis of bacterial pathogens.
- This chip-based approach has potential for rapid and sensitive pathogen detection systems.
