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Rapid and sensitive immunodetection of Listeria monocytogenes in milk using a novel piezoelectric cantilever sensor
1Department of Chemical & Biological Engineering, Drexel University, Philadelphia, PA 19104, USA.
Abstract:
Listeria monocytogenes (LM), an important food-borne pathogen that has a high mortality rate (≈ 30%), was successfully detected within an hour at the infection dose limit of 10(3)/mL, both in buffer and milk. LM detection was demonstrated using a novel asymmetrically anchored cantilever sensor and a commercially available antibody. Sensor responses were confirmed using a secondary antibody binding step, similar to the sandwich ELISA assays, as a means of signal amplification that also reduced the occurrence of false negatives. Detection of LM at a concentrations of 10(2)/mL was achieved, by incorporating a third antibody binding step, which is an order of magnitude smaller than the infection dose (<1000 cells) for LM. The commercially available antibody for LM used in this work is shown to have low avidity which partially explains the relatively low sensitivity reported for LM as compared to other pathogens.
Insights
This study presents a novel sensor for rapid Listeria monocytogenes (LM) detection in food and clinical samples. The sensor achieved detection at clinically relevant levels, offering a promising tool for foodborne pathogen surveillance.
Area of Science:
- Food safety
- Microbiology
- Biosensor technology
Background:
- Listeria monocytogenes (LM) is a significant foodborne pathogen with a high mortality rate.
- Rapid and sensitive detection methods are crucial for preventing LM outbreaks.
Purpose of the Study:
- To develop and validate a novel asymmetrically anchored cantilever sensor for LM detection.
- To assess the sensor's performance in buffer and milk matrices.
Main Methods:
- Utilized a novel asymmetrically anchored cantilever sensor with a commercially available antibody.
- Employed secondary and tertiary antibody binding steps for signal amplification and enhanced sensitivity, mimicking sandwich ELISA principles.
Main Results:
- Successfully detected LM within one hour at an infection dose limit of 10(3) cells/mL in buffer and milk.
- Achieved detection at concentrations as low as 10(2) cells/mL with a three-antibody approach.
- Identified low antibody avidity as a factor influencing LM detection sensitivity.
Conclusions:
- The developed cantilever sensor demonstrates rapid and sensitive detection of Listeria monocytogenes.
- The sensor shows potential for application in food safety and clinical diagnostics.
- Further optimization, potentially involving higher avidity antibodies, could improve detection limits.
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