Related Experiment Video
Updated: Jul 29, 2026
Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres
Published on: October 23, 2011
The Detection of Foodborne Pathogenic Bacteria in Seafood Using a Multiplex Polymerase Chain Reaction System
Pengzhen Li1, Xiaoxuan Feng1, Baiyan Chen1
1Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University, Guangzhou 510642, China.
Abstract:
Multiplex polymerase chain reaction (PCR) assays are mainly used to simultaneously detect or identify multiple pathogenic microorganisms. To achieve high specificity for detecting foodborne pathogenic bacteria, specific primers need to be designed for the target strains. In this study, we designed and achieved a multiplex PCR system for detecting eight foodborne pathogenic bacteria using specific genes: toxS for Vibrio parahaemolyticus, virR for Listeria monocytogenes, recN for Cronobacter sakazakii, ipaH for Shigella flexneri, CarA for Pseudomonas putida, rfbE for Escherichia coli, vvhA for Vibrio vulnificus, and gyrB for Vibrio alginolyticus. The sensitivity of the single system in this study was found to be 20, 1.5, 15, 15, 13, 14, 17, and 1.8 pg for V. parahaemolyticus, L. monocytogenes, E. coli O157:H7, C. sakazakii, S. flexneri, P. putida, V. vulnificus, and V. alginolyticus, respectively. The minimum detection limit of the multiplex system reaches pg/μL detection level; in addition, the multiplex system exhibited good specificity and stability. Finally, the assays maintained good specificity and sensitivity of 104 CFU/mL for most of the samples and we used 176 samples of eight aquatic foods, which were artificially contaminated to simulate the detection of real samples. In conclusion, the multiplex PCR method is stable, specific, sensitive, and time-efficient. Moreover, the method is well suited for contamination detection in these eight aquatic foods and can rapidly detect pathogenic microorganisms.
Insights
This study developed a multiplex polymerase chain reaction (PCR) assay for detecting eight common foodborne pathogens. The new method offers a rapid, stable, and sensitive approach for food safety testing.
Area of Science:
- Microbiology
- Molecular Biology
- Food Science
Background:
- Multiplex polymerase chain reaction (PCR) assays are crucial for simultaneously detecting multiple pathogenic microorganisms.
- High specificity in detecting foodborne pathogenic bacteria requires the design of specific primers for target strains.
Purpose of the Study:
- To design and validate a multiplex PCR system for the simultaneous detection of eight foodborne pathogenic bacteria.
- To assess the specificity, sensitivity, and stability of the developed multiplex PCR assay.
Main Methods:
- Designed specific primers targeting unique genes (toxS, virR, recN, ipaH, CarA, rfbE, vvhA, gyrB) for eight foodborne pathogens: Vibrio parahaemolyticus, Listeria monocytogenes, Cronobacter sakazakii, Shigella flexneri, Pseudomonas putida, Escherichia coli, Vibrio vulnificus, and Vibrio alginolyticus.
- Determined the sensitivity of single and multiplex systems, with minimum detection limits reaching the pg/μL level.
- Evaluated specificity and stability using 176 artificially contaminated aquatic food samples.
Main Results:
- The multiplex PCR system demonstrated high specificity and stability.
- Sensitivity reached pg/μL levels for individual pathogen detection.
- The assay maintained good specificity and sensitivity (10^4 CFU/mL) in contaminated aquatic food samples.
Conclusions:
- The developed multiplex PCR method is stable, specific, sensitive, and time-efficient.
- This method is well-suited for rapid contamination detection in aquatic foods.
- The assay effectively detects pathogenic microorganisms in food samples.
More Related Videos
06:55A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.
Published on: November 7, 2018
10:33Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis
Published on: June 17, 2019
