Simultaneous detection of Salmonella spp., Pseudomonas aeruginosa, Bacillus cereus, and Escherichia coli O157:H7 in environmental water using PMA combined with mPCR

Guoyang Xie; Shuang Yu; Wen Li; Dan Mu; Zoraida P. Aguilar; Hengyi Xu

doi:10.1007/s12275-020-0084-6

Articles

Page Path: HOME > J. Microbiol > Volume 58(8); 2020 > Article

Journal Article
Simultaneous detection of Salmonella spp., Pseudomonas aeruginosa, Bacillus cereus, and Escherichia coli O157:H7 in environmental water using PMA combined with mPCR: Guoyang Xie ¹, Shuang Yu ¹, Wen Li ¹, Dan Mu ¹, Zoraida P. Aguilar ², Hengyi Xu ¹; Journal of Microbiology 2020;58(8):668-674.
DOI: https://doi.org/10.1007/s12275-020-0084-6
Published online: June 25, 2020

¹State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, P. R. China, ²Zystein, LLC., Fayetteville, Arkansas 72703, USA¹State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, P. R. China, ²Zystein, LLC., Fayetteville, Arkansas 72703, USA

Corresponding author: Hengyi Xu , Tel: +86-791-8830-4447-ext-9520,

Received: 17 February 2020 • Revised: 18 May 2020 • Accepted: 20 May 2020

prev next

50 Views
0 Download
12 Web of Science
10 Crossref
15 Scopus

Full Article

Abstract

A multiplex polymerase chain reaction (mPCR) with propidium monoazide (PMA) and internal amplification control (IAC) for the simultaneous detection of waterborne pathogens Salmonella spp., Pseudomonas aeruginosa, Bacillus cereus, and Escherichia coli O157:H7, was developed. This PMA-IAC-mPCR assay used four new specific primers based on the genes for invA, ecfX, cesB, and fliC, respectively. A 16S rRNA primer was chosen for IAC to eliminate false negative
results
. The photosensitive dye, propidium monoazide (PMA) was used to exclude signals from dead bacteria that could lead to false positive results. In pure culture, the limits of detection (LOD) were 101 CFU/ml for P. aeruginosa, 102 CFU/ml for both Salmonella spp. and E. coli O157:H7, and 103 CFU/ml for B. cereus, respectively. In addition, with a 6–8 h enrichment of all four bacteria that were combined in a mixture that was spiked in water sample matrix, the LOD was 3 CFU/ml for Salmonella spp., 7 CFU/ml for E. coli O157:H7, 10 CFU/ml for B. cereus and 2 CFU/ml for P. aeruginosa. This PMA-IAC-mPCR assay holds potential for application in the multiplex assay of waterborne pathogens.

Keywords: SARS-CoV-2;monoclonal antibodies;neutralization;transmission;vaccine;variants

Figure & Data

References

Citations

Citations to this article as recorded by

Rapid, sensitive, and user-friendly detection of Pseudomonas aeruginosa using the RPA/CRISPR/Cas12a system
Wenjing Zhang, Hai Qu, Xin Wu, Jingjing Shi, Xinling Wang
BMC Infectious Diseases.2024;[Epub] CrossRef
Review of Detection Limits for Various Techniques for Bacterial Detection in Food Samples
Xinyi Zhao, Abhijnan Bhat, Christine O’Connor, James Curtin, Baljit Singh, Furong Tian
Nanomaterials.2024; 14(10): 855. CrossRef
The dual nucleic acid amplification with dynamic light scattering strategy for ultrasensitive detection of Salmonella in milk
Qian Xu, Guoyang Xie, Qiang Shi, Ju Liu, Baoqing Zhou, Ping Tong, Zoraida P. Aguilar, Hengyi Xu
Microchemical Journal.2023; 184: 108143. CrossRef
An Assay Combining Droplet Digital PCR With Propidium Monoazide Treatment for the Accurate Detection of Live Cells of Vibrio vulnificus in Plasma Samples
Ling Hu, Yidong Fu, Shun Zhang, Zhilei Pan, Jiang Xia, Peng Zhu, Jing Guo
Frontiers in Microbiology.2022;[Epub] CrossRef
Molecular Diagnostic Tools Applied for Assessing Microbial Water Quality
Lisa Paruch
International Journal of Environmental Research and Public Health.2022; 19(9): 5128. CrossRef
Rapid-Response Magnetic Enrichment Strategy for Significantly Improving Sensitivity of Multiplex PCR Analysis of Pathogenic Listeria Species
Fangbin Xiao, Xuekun Bai, Keyu Wang, Yifan Sun, Hengyi Xu
Applied Sciences.2022; 12(13): 6415. CrossRef
Real-time and visual detection of viableSalmonellain milk by a competitive annealing mediated isothermal amplification (CAMP) combined with propidium monoazide (PMA)
Xu Chen, Wei Li, Yue Ma
Analytical Methods.2022; 14(38): 3773. CrossRef
Hybrid RCA-DLS assay combined with aPCR for sensitive Salmonella enteritidis detection
Guoyang Xie, Zhongxu Zhan, Yu Ye, Baoqing Zhou, Ping Tong, Zoraida P. Aguilar, Hengyi Xu
Analytical Biochemistry.2022; 646: 114647. CrossRef
How to Evaluate Non-Growing Cells—Current Strategies for Determining Antimicrobial Resistance of VBNC Bacteria
Susanne Fleischmann, Christian Robben, Thomas Alter, Peter Rossmanith, Patrick Mester
Antibiotics.2021; 10(2): 115. CrossRef
Development of a simple, rapid multiplex PCR tool kit by using the 16S rRNA gene for the identification of faecal and non-faecal coliforms in drinking water
A. Shiva Shanker, N. Rajesh, Pavan Kumar Pindi
Water Supply.2021; 21(7): 3319. CrossRef

Cite this Article

Cite this Article: export Copy Download Format; Close

Download Citation

Download a citation file in RIS format that can be imported by all major citation management software, including EndNote, ProCite, RefWorks, and Reference Manager.

Format:

RIS — For EndNote, ProCite, RefWorks, and most other reference management software
BibTeX — For JabRef, BibDesk, and other BibTeX-specific software

Include:

Citation for the content below
Citation and abstract for the content below

Simultaneous detection of Salmonella spp., Pseudomonas aeruginosa, Bacillus cereus, and Escherichia coli O157:H7 in environmental water using PMA combined with mPCR

J. Microbiol. 2020;58(8):668-674. Published online June 25, 2020

DOI: https://doi.org/10.1007/s12275-020-0084-6

Related articles