Application of a multiplex PCR for the simultaneous detection of Escherichia coli O157:H7, Salmonella and Shigella in raw and ready-to-eat meat products

Meat Science ◽  
2005 ◽  
Vol 71 (2) ◽  
pp. 402-406 ◽  
Author(s):  
Y. Li ◽  
S. Zhuang ◽  
A. Mustapha
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Meat Products ◽  
Escherichia Coli O157

A gold nanoparticles-assisted multiplex PCR assay for simultaneous detection of Salmonella typhimurium, Listeria monocytogenes and Escherichia coli O157:H7

2020 ◽  
Vol 12 (2) ◽  
pp. 212-217 ◽  
Author(s):  
Juan Du ◽  
Shujing Wu ◽  
Liyuan Niu ◽  
Junguang Li ◽  
Dianbo Zhao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gold Nanoparticles ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Escherichia Coli O157 ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Pcr Product

Unfunctionalized flower-shaped AuNPs is used as colorimetric sensor for PCR product detection by naked eyes.

Multiplex PCR for Simultaneous Detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 in Meat Samples

2005 ◽  
Vol 68 (3) ◽  
pp. 551-556 ◽  
Author(s):  
SUSUMU KAWASAKI ◽  
NAOKO HORIKOSHI ◽  
YUKIO OKADA ◽  
KAZUKO TAKESHITA ◽  
TAKASHI SAMESHIMA ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Detection Sensitivity ◽  
Rapid Screening ◽  
Escherichia Coli O157 ◽  
Salmonella Spp ◽  
E Coli ◽  
Meat Samples

A multiplex PCR method was developed for simultaneous detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 in meat samples. DNA detection sensitivity for this method was 103 CFU/ml for each pathogen. When this protocol was used for the detection of each of the above pathogenic bacteria in spiked pork samples, 1 cell per 25 g of inoculated sample could be detected within 30 h. In the samples of naturally contaminated meat, Salmonella spp., L. monocytogenes, and E. coli O157:H7 were detected over the same time period. Excellent agreement was obtained for the results of multiplex PCR and the conventional culture method, which suggests that the multiplex PCR is a reliable and useful method for rapid screening of meat products for Salmonella spp., L. monocytogenes, and E. coli O157:H7 contamination.

scholarly journals Reverse Transcription-Multiplex PCR Assay for Simultaneous Detection of Escherichia coli O157:H7, Vibrio cholerae O1, and Salmonella Typhi

2004 ◽  
Vol 50 (11) ◽  
pp. 2037-2044 ◽  
Author(s):  
Nicole J Morin ◽  
Zhilong Gong ◽  
Xing-Fang Li
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Vibrio Cholerae ◽  
Reverse Transcription ◽  
Simultaneous Detection ◽  
Salmonella Typhi ◽  
Escherichia Coli O157 ◽  
Single Tube ◽  
E Coli ◽  
Vibrio Cholerae O1

Abstract Background: Escherichia coli O157:H7, Vibrio cholerae O1, and Salmonella Typhi are pathogenic bacteria that can be found in contaminated water supplies throughout the world. No currently available assays can simultaneously detect and identify all three pathogens. Our aim was to develop a rapid and reliable technique for simultaneous detection of these pathogens. Methods: Four unique genes were chosen as the targets of detection. Forward and reverse primers were designed to specifically amplify different sizes of these target genes: a 239-bp region of the E. coli O157 lipopolysaccharide (LPS) gene (rfbE); a 179-bp region of the H7 flagellin gene (fliC); a 419-bp region of the V. cholerae O1 LPS gene (rfbE); and a 329-bp region of Salmonella Typhi LPS gene (tyv). To ensure the detection of only viable replicating bacteria, RNA was extracted for analysis. After reverse transcription, cDNAs were simultaneously amplified in a single tube by multiplex PCR. The multiplex PCR products were analyzed by gel electrophoresis. To characterize the assay we analyzed, in a blinded fashion, seven unknown RNA samples containing various combinations of total RNA from these bacteria as well as clinical isolates. Results: All seven unknown RNA samples were correctly identified. The assay was able to detect and identify as few as 30 cells of E. coli O157:H7 and Salmonella Typhi in clinical isolates, and the presence of other bacteria did not interfere with the analysis. Conclusion: An assay combining reverse transcription with single-tube multiplex PCR was successfully developed and validated for simultaneous detection of viable E. coli O157:H7, V. cholerae O1, and Salmonella Typhi.

A Novel Multiplex PCR Assay for Rapid and Simultaneous Detection of Five Pathogenic Bacteria: Escherichia coli O157:H7, Salmonella, Staphylococcus aureus, Listeria monocytogenes, and Vibrio parahaemolyticus

2007 ◽  
Vol 70 (7) ◽  
pp. 1656-1662 ◽  
Author(s):  
JEONG SOON KIM ◽  
GANG GWEON LEE ◽  
JONG SEOK PARK ◽  
YONG HYUN JUNG ◽  
HYO SUN KWAK ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Multiplex Pcr ◽  
Vibrio Parahaemolyticus ◽  
Pathogenic Bacteria ◽  
Simultaneous Detection ◽  
Escherichia Coli O157 ◽  
Pcr Assay ◽  
Foodborne Pathogenic Bacteria ◽  
Single Reaction

Rapid and sensitive detection techniques for foodborne pathogens are important to the food industry. However, traditional detection methods rely on bacterial culture in combination with biochemical tests, a process that typically takes 4 to 7 days to complete. Thus, this study was conducted to address the issue of time lag inherent in traditional methods by developing a novel PCR assay for each of five foodborne pathogenic bacteria. This new system consists of a simultaneous screening method using multiplex PCR in a single reaction tube for the rapid and sensitive detection of each of the five bacteria. Specific primers for multiplex PCR amplification of the Shiga-like toxin (verotoxin type II), femA (cytoplasmic protein), toxR (transmembrane DNA binding protein), iap (invasive associative protein), and invA (invasion protein A) genes were designed to allow simultaneous detection of Escherichia coli O157:H7, Staphylococcus aureus, Vibrio parahaemolyticus, Listeria monocytogenes, and Salmonella, respectively. To confirm the specificity of each primer pair for the respective target gene, three types of experiments were carried out using boiled cell lysates and their DNAs. In the multiplex PCR with mixed DNA samples, specific bands for corresponding genes were simultaneously detected from a single reaction. The detection of all five foodborne pathogenic bacteria could be completed in less than 24 h with this novel PCR method.

scholarly journals Evaluation of Multiplex PCR System for Simultaneous Detection of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enteritidis in Shrimp Samples

1970 ◽  
Vol 24 (1) ◽  
pp. 42-46 ◽  
Author(s):  
Mahmuda Yasmin ◽  
Susumu Kawasaki ◽  
Shinichi Kawamoto
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Multiplex Pcr ◽  
Salmonella Enteritidis ◽  
Simultaneous Detection ◽  
Dna Amplification ◽  
Rapid Screening ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Pcr Method

A multiplex polymerase chain reaction (PCR) method was evaluated for simultaneous detection of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enteritidis in shrimp samples. The sensitivity of DNA amplification by PCR in this method was found to be 103 cfu/ml for each pathogen. When this protocol was adopted for the detection of each of the above mentioned pathogen in spiked shrimp extract culture, similar sensitivity was observed. However, this method detected 1 bacterial cell for E. coli O157:H7 and S. enteritidis and 100 for L. monocytogenes per 25 g spiked shrimp samples after overnight enrichment. In the commercially imported shrimp samples, none was found to contain any of the three pathogens by multiplex PCR or by conventional method, which suggests that the multiplex PCR is a reliable and useful for rapid screening of shrimp samples for E. coli O157:H7, L. monocytogenes and S. enteritidis. This will save time and increase our ability to assure food safety. Keywords: Multiples PCR, Shrimp extract, Spiked sample, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enteritidisDOI: http://dx.doi.org/10.3329/bjm.v24i1.1236 Bangladesh J Microbiol, Volume 24, Number 1, June 2007, pp 42-46

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