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Multiplex quantitative foodborne pathogen detection using high resolution CE-SSCP coupled stuffer-free multiplex
Boram Chung1, Gi Won Shin, Jeongkyeong Na
1School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Korea.
A new stuffer-free multiplex ligation-dependent probe amplification (MLPA) method combined with capillary electrophoresis-single-strand conformation polymorphism (CE-SSCP) enables sensitive detection of ten foodborne pathogens. This technique offers a reliable tool for enhancing food safety through precise pathogen identification.
Area of Science:
- Microbiology
- Molecular Biology
- Food Safety Science
Background:
- Sensitive detection of foodborne pathogens is crucial for food safety.
- Genetic marker detection is a key strategy for sensitive pathogen identification.
- Existing methods like capillary electrophoresis (CE) have limitations in multiplex detection of specific markers.
Purpose of the Study:
- To develop a sensitive and reliable method for detecting multiple foodborne pathogens.
- To overcome limitations of existing multiplex ligation-dependent probe amplification (MLPA) methods for foodborne pathogen detection.
Main Methods:
- Development of a stuffer-free MLPA technique.
- Integration of MLPA with high-resolution capillary electrophoresis-single-strand conformation polymorphism (CE-SSCP).
- Design of species-specific probes for targeted pathogen detection.
Main Results:
- The developed MLPA-CE-SSCP method successfully identified ten foodborne pathogens.
- Achieved limits of detection ranging from 0.5-5 pg of genomic DNA.
- Demonstrated quantitative determination of over 100-fold increases in pathogen DNA.
Conclusions:
- Stuffer-free MLPA combined with CE-SSCP is a sensitive and reliable technique for foodborne pathogen detection.
- This method enhances the capability for precise identification of multiple pathogens.
- The technique holds significant potential for improving food safety monitoring.
