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Phage M13KO7 detection with biosensor based on imaging ellipsometry and AFM microscopic confirmation
1Institute of Mechanics, Chinese Academy of Sciences, Beijing 100190, China.
Abstract:
A rapid detection and identification of pathogens is important for minimizing transfer and spread of disease. A label-free and multiplex biosensor based on imaging ellipsometry (BIE) had been developed for the detection of phage M13KO7. The surface of silicon wafer is modified with aldehyde, and proteins can be patterned homogeneously and simultaneously on the surface of silicon wafer in an array format by a microfluidic system. Avidin is immobilized on the surface for biotin-anti-M13 immobilization by means of interaction between avidin and biotin, which will serve as ligand against phage M13KO7. Phages M13KO7 are specifically captured by the ligand when phage M13KO7 solution passes over the surface, resulting in a significant increase of mass surface concentration of the anti-M13 binding phage M13KO7 layer, which could be detected by imaging ellipsometry with a sensitivity of 10(9)pfu/ml. Moreover, atomic force microscopy is also used to confirm the fact that phage M13KO7 has been directly captured by ligands on the surface. It indicates that BIE is competent for direct detection of phage M13KO7 and has potential in the field of virus detection.
Insights
A novel biosensor uses imaging ellipsometry for rapid, label-free detection of the M13KO7 bacteriophage. This advancement offers sensitive virus detection, crucial for disease control and pathogen identification.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Microfluidics
Background:
- Rapid pathogen detection is vital for disease control.
- Existing methods can be time-consuming or lack multiplexing capabilities.
- Biosensors offer a promising avenue for sensitive and specific pathogen identification.
Purpose of the Study:
- To develop and validate a label-free, multiplex biosensor for detecting bacteriophage M13KO7.
- To assess the sensitivity and specificity of the biosensor system.
- To demonstrate the potential of imaging ellipsometry in virus detection.
Main Methods:
- Development of a biosensor utilizing imaging ellipsometry (BIE).
- Surface modification of silicon wafers with aldehyde and homogeneous protein patterning using microfluidics.
- Immobilization of avidin for biotin-anti-M13 binding as a ligand for M13KO7 capture.
- Detection of captured M13KO7 via mass surface concentration changes measured by BIE.
- Confirmation of phage capture using atomic force microscopy.
Main Results:
- The BIE biosensor achieved label-free and multiplex detection of M13KO7.
- A sensitivity of 10^9 plaque-forming units/ml was demonstrated for M13KO7 detection.
- Atomic force microscopy confirmed direct capture of M13KO7 by the immobilized ligands.
- The system showed specific capture of M13KO7 when the phage solution passed over the surface.
Conclusions:
- The developed BIE biosensor is effective for the direct and rapid detection of M13KO7.
- Imaging ellipsometry demonstrates significant potential for sensitive virus detection applications.
- This technology could aid in minimizing disease spread through early pathogen identification.
