[Development of multiplex oligonucleotide ligation-PCR-universal DNA microarrays for detection of foodborne...

Area of Science:

• Microbiology
• Molecular Biology
• Biotechnology

Background:

• Foodborne bacterial infections pose significant public health risks.
• Accurate and rapid detection of foodborne pathogens is crucial for food safety.
• Existing detection methods can be time-consuming or lack multiplexing capabilities.

Purpose of the Study:

• To develop a novel multiplex oligonucleotide ligation-polymerase chain reaction (MOL-PCR) based universal microarray assay.
• To achieve multiplexed detection of common foodborne pathogens.
• To establish a rapid, accurate, and high-throughput detection platform.

Main Methods:

• Design of specific upstream and downstream probes for eight common foodborne pathogens.
• Enrichment of target fragments via multiplex PCR.
• Generation of fluorescently labeled amplicons using multiplex ligase detection reaction and asymmetric PCR.
• Detection of products via hybridization to immobilized tag sequences on DNA microarrays.

Main Results:

• The assay successfully identified all eight target pathogens in single and mixed infections.
• Achieved a detection limit of (1.1–8.5) × 10^2 CFU/mL for pure bacterial cultures.
• Microarray results showed high concordance with traditional culture, biochemical identification, and real-time PCR for 96 food and clinical samples.

Conclusions:

• The developed MOL-PCR universal microarray assay is a novel platform for foodborne pathogen detection.
• The assay offers rapid, accurate, sensitive, and high-throughput identification of pathogenic bacteria.
• This method has significant potential for improving food safety monitoring and public health.