Fast and sensitive detection of foodborne pathogen using electrochemical impedance analysis, urease catalysis and microfluidics

• Published in: Biosensors & bioelectronics 2016-12, Vol.86, p.770-776
• Main Authors: Chen, Qi, Wang, Dan, Cai, Gaozhe, Xiong, Yonghua, Li, Yuntao, Wang, Maohua, Huo, Huiling, Lin, Jianhan
• Format: Article
• Language: English
• Subjects: Bacteria; Bacterial Typing Techniques - instrumentation; Biosensors; Catalysis; Chips; Dielectric Spectroscopy - instrumentation; Equipment Design; Equipment Failure Analysis; Food Analysis - instrumentation; Food Contamination - analysis; Food Microbiology - instrumentation; Immunoassay - instrumentation; Immunomagnetic separation; Lab-On-A-Chip Devices; Listeria; Listeria - immunology; Listeria - isolation & purification; Listeria monocytogenes; Microfluidic impedance biosensor; Microfluidics; Nanoparticles; Phase angle; Reproducibility of Results; Sensitivity and Specificity; Separation; Urease - chemistry; Urease - immunology; Urease catalysis
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2016.07.071

Early screening of pathogenic bacteria is a key to prevent and control of foodborne diseases. In this study, we developed a fast and sensitive bacteria detection method integrating electrochemical impedance analysis, urease catalysis with microfluidics and using Listeria as model. The Listeria cells, the anti-Listeria monoclonal antibodies modified magnetic nanoparticles (MNPs), and the anti-Listeria polyclonal antibodies and urease modified gold nanoparticles (AuNPs) were incubated in a fluidic separation chip with active mixing to form the MNP-Listeria-AuNP-urease sandwich complexes. The complexes were captured in the separation chip by applying a high gradient magnetic field, and the urea was injected to resuspend the complexes and hydrolyzed under the catalysis of the urease on the complexes into ammonium ions and carbonate ions, which were transported into a microfluidic detection chip with an interdigitated microelectrode for impedance measurement to determine the amount of the Listeria cells. The capture efficiency of the Listeria cells in the separation chip was ∼93% with a shorter time of 30min due to the faster immuno-reaction using the active magnetic mixing. The changes on both impedance magnitude and phase angle were demonstrated to be able to detect the Listeria cells as low as 1.6×102CFU/mL. The detection time was reduced from original ∼2h to current ∼1h. The recoveries of the spiked lettuce samples ranged from 82.1% to 89.6%, indicating the applicability of this proposed biosensor. This microfluidic impedance biosensor has shown the potential for online, automatic and sensitive bacteria separation and detection. •Phase angle shift was successfully used for determination of Listeria.•The detection limit of this biosensor was 102CFU/mL for Listeria.•The detection time was reduced to 1h due to on-chip passive mixing.•The recoveries of Listeria cells in the spiked samples was ∼85%.