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Detection of Neisseria meningitidis and Yersinia pestis with a novel silicon-based sensor
Abstract:
A light-addressable potentiometric (silicon) sensor was used in an immunofiltration procedure for the detection of pathogenic bacteria. Yersinia pestis was detected by filtering the cells onto nitrocellulose membranes and then filtering anti-Y. pestis mouse monoclonal antibody and anti-mouse immunoglobulin G-horseradish peroxidase conjugate. For Neisseria meningitidis detection, mouse monoclonal antibody to the major outer membrane protein of this bacterium was coupled directly to horseradish peroxidase. N. meningitidis cell suspensions were filtered onto polycarbonate membranes, and the enzyme conjugate was allowed to react with the filtered bacteria. The presence of both enzyme conjugates was determined potentiometrically with the silicon sensor. The sensitivity of this technique relative to that of an enzyme-linked immunosorbent assay for N. meningitidis was determined. Fewer than 1,000 bacterial cells could be detected with the silicon sensor in a 20-min assay, whereas a 2.5-h enzyme-linked immunosorbent assay with the same antigen and antibody preparations was significantly less sensitive.
Insights
A novel immunofiltration assay using a light-addressable potentiometric sensor detects pathogenic bacteria like Yersinia pestis and Neisseria meningitidis with high sensitivity. This method offers a faster and more sensitive alternative to traditional enzyme-linked immunosorbent assays.
Area of Science:
- Biomedical Engineering
- Microbiology
- Sensor Technology
Background:
- Pathogenic bacteria detection is crucial for public health.
- Current methods like ELISA can be time-consuming and less sensitive.
- Need for rapid and sensitive diagnostic tools for bacterial infections.
Purpose of the Study:
- To develop and evaluate a light-addressable potentiometric sensor-based immunofiltration assay for detecting pathogenic bacteria.
- To compare the sensitivity and speed of this new method against enzyme-linked immunosorbent assay (ELISA).
Main Methods:
- Utilized a light-addressable potentiometric sensor in an immunofiltration setup.
- Detected Yersinia pestis using filtration of cells, antibodies, and enzyme conjugates.
- Detected Neisseria meningitidis by filtering cells and reacting with enzyme-coupled antibodies.
- Quantified enzyme conjugates potentiometrically with the silicon sensor.
Main Results:
- Successfully detected both Yersinia pestis and Neisseria meningitidis.
- The silicon sensor assay detected fewer than 1,000 bacterial cells.
- Achieved detection in a rapid 20-minute assay time.
- Demonstrated significantly higher sensitivity compared to a 2.5-hour ELISA.
Conclusions:
- The light-addressable potentiometric sensor-based immunofiltration assay is a highly sensitive and rapid method for detecting pathogenic bacteria.
- This technique offers a significant advantage over conventional ELISA in terms of speed and sensitivity.
- Potential for application in clinical diagnostics and food safety testing.