Rapid and Simultaneous Detection of Five, Viable, Foodborne Pathogenic Bacteria by Photoinduced PMAxx-Coupled Multiplex PCR in Fresh Juice

2021 ◽  
Author(s):  
Tiantian Huang ◽  
Yaoqiang Shi ◽  
Jinyang Zhang ◽  
Qinqin Han ◽  
Xue-shan Xia ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Pathogenic Bacteria ◽  
Simultaneous Detection ◽  
Foodborne Pathogenic Bacteria

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 Multiplex PCR Assay for the Simultaneous Detection of Major Pathogenic Bacteria in Soybean

2013 ◽  
Vol 58 (2) ◽  
pp. 142-148 ◽  
Author(s):  
Yeong-Hoon Lee ◽  
Nam-Goo Kim ◽  
Young-Nam Yoon ◽  
Seung-Taek Lim ◽  
Hyun-Tae Kim ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Pathogenic Bacteria ◽  
Simultaneous Detection ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

scholarly journals A multiplex PCR assay for simultaneous detection of four major pathogenic bacteria of ayu

2019 ◽  
Vol 85 (3) ◽  
pp. 340-342
Author(s):  
KYUMA SUZUKI ◽  
SHOTARO IZUMI ◽  
SHINJI KUMAKAWA ◽  
TAKAMITU SAKAI ◽  
CHIHAYA NAKAYASU
Keyword(s):  
Multiplex Pcr ◽  
Pathogenic Bacteria ◽  
Simultaneous Detection ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

Multiplex PCR method for simultaneous detection of five pathogenic bacteria closely related to foodborne diseases

3 Biotech ◽  
2021 ◽  
Vol 11 (5) ◽  
Author(s):  
Ying Chen ◽  
Zixuan Wang ◽  
Qiaozhen Shi ◽  
Shengxiong Huang ◽  
Taotao Yu ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Pathogenic Bacteria ◽  
Simultaneous Detection ◽  
Foodborne Diseases ◽  
Pcr Method

Rapid and Simultaneous Detection of Vibrio parahaemolyticus, Salmonella spp. and Escherichia coli in Fish by Multiplex PCR

2020 ◽  
Vol 15 (2) ◽  
pp. 53
Author(s):  
Radestya Triwibowo ◽  
Novalia Rachmawati ◽  
Dwiyitno Dwiyitno
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Internal Control ◽  
Vibrio Parahaemolyticus ◽  
Pathogenic Bacteria ◽  
Target Genes ◽  
Simultaneous Detection ◽  
Salmonella Spp ◽  
Fish Products ◽  
Confirmatory Assay

Pathogenic bacteria are commonly found as natural contaminants in seafood and fish products. Globally, several countries have been imposing strict regulations on the maximum levels of pathogens and consequently require microbial testing of pathogens before the products can be marketed. A culture-based method with biochemical assay has been widely used to detect pathogenic bacteria in food, despite its long and extensive process. Meanwhile, the alternative molecular-based method to overcome this problem, cannot differentiate between viable and nonviable cells, which may lead to underestimation. This study aimed to develop a multiplex PCR (mPCR) method as a confirmatory assay for the culture-based method to detect pathogens in fish products simultaneously. This method applied a pre-enrichment step to ensure the growth of low-level pathogens and the injured cells in the sample. The target genes were ToxR, InvA, and UidA for Vibrio parahaemolyticus, Salmonella spp. and Escherichia coli, respectively. This assay also amplified the 16S rDNA gene of bacteria as an internal control for the PCR reaction. By implementing liquid-based DNA extraction during analysis, the developed-mPCR was comparable to detect the targeted bacteria in artificially-contaminated samples. The method was more sensitive in naturally-contaminated samples, where the number of E. coli, Salmonella spp. and V. parahaemolyticus detected were 28, 7, and 22, respectively. While the conventional method only detected 26, 5, and 19 of the respective pathogens. With a relatively shorter time and lower operation cost, the mPCR method is potential as an alternative for the culture-based method.

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