4.3 Article

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

Journal

3 BIOTECH
Volume 11, Issue 5, Pages -

Publisher

SPRINGER HEIDELBERG
DOI: 10.1007/s13205-021-02759-y

Keywords

Multiplex PCR; S; aureus; L; monocytogenes; S; flexneri; Y; enterocolitica; C; difficile; Detection method; Foodborne Diseases

Funding

  1. National Natural Science Foundation of China [81702103]
  2. Jiangsu Provincial Natural Science Foundation [BK20170252]
  3. Projects for Jiangsu Provincial Young Medical Talents [QNRC2016780]

Ask authors/readers for more resources

This study presents a multiplex PCR method for detecting five food-relevant virulence pathogenicity genes of intestinal pathogens, offering a rapid and accurate approach for pathogen identification.
In this study, we describe a multiplex PCR method for the detection of five food-relevant virulence pathogenicity genes of intestinal pathogens. Five pairs of primers were designed based on nuc gene for Staphylococcus aureus, hlyA gene of Listeria monocytogenes, ipaH gene of Shigella flexneri, lysP gene of Yersinia enterocolitica and tpi gene of Clostridium difficile. Conditions were optimized to amplify fragments of those genes simultaneously in one PCR amplification. After developing and optimizing the multiplex PCR reaction system, the specificity and sensitivity of the multiple PCR assays were evaluated. The optimized program is also applied to retail meat for testing. The result indicated that when the annealing temperature was 54 degrees C and the primer concentrations of S. aureus, L. monocytogenes, S. flexneri, Y. enterocolitica and C. difficile are 10, 10, 5, 3 and 2 mu M, the five strains could expand 484, 345, 204, 156, 88 bp of clear fragments, respectively. So was the multiple PCR in artificially contaminated beef produce. All cultures were cultured and separated by traditional methods. The multiplex PCR method offers a rapid, simple, and accurate identification of pathogens and could be used in food safety investigations, clinical diagnosis as well as for the surveillance of the spreading determinants of pathogens in epidemiological studies.

Authors

I am an author on this paper
Click your name to claim this paper and add it to your profile.

Reviews

Primary Rating

4.3
Not enough ratings

Secondary Ratings

Novelty
-
Significance
-
Scientific rigor
-
Rate this paper

Recommended

No Data Available
No Data Available