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Updated: Aug 23, 2026
Detection of Live Escherichia coli O157:H7 Cells by PMA-qPCR
Published on: February 1, 2014
A study on detecting and identifying enteric pathogens with PCR
Jun-Wen Li1, Xiu-Quan Shi, Fu-Huan Chao
1Department of Environment and Health, Institute of Health and Environmental Medicine, 1 Da Li Road, Tianjin 300050, China. junwenli@eyou.com
Objective:
To develop a rapid and definite diagnostic test of bacterial enteritis caused by pathogenic enterobacteria, the most frequent etiologic agent of infectious enteritis in the world.
Methods:
A set of conventional PCR assays were applied to detect and identify salmonella, shigella, and E. coli O157:H7 directly from pure culture and fecal samples. The general primers of pathogenic enterobacteria were located on the uidA gene, which were found not only in E. coli nuclear acid, but also in shigella and salmonella genes. Shigella primer was from ipaH gene whose coded invasive plasmid relative antigen existed both in plasmid and in genome. The primers of salmonella were designed from the 16SrRNA sequence. The primer of E. coli O157:H7 was taken from eaeA gene. Five random primers were selected for RAPD. The detection system included common PCR, semi-nested PCR and RAPD.
Results:
This method was more sensitive, specific and efficient and its processing was rapid and simple. For example, the method could be used to specifically detect and identify salmonella, shigella, and E. coli O157:H7, and its sensitivity ranged from 3 to 50 CFU, and its detection time was 4 hours.
Conclusion:
This PCR method, therefore, can serve as a routine and practical protocol for detecting and identifying pathogenic microorganisms from clinical samples.
Insights
A new PCR method rapidly and accurately detects bacterial enteritis pathogens like Salmonella, Shigella, and E. coli O157:H7. This sensitive test identifies as few as 3-50 CFU in just 4 hours, aiding clinical diagnosis.
Area of Science:
- Microbiology
- Molecular Biology
- Clinical Diagnostics
Background:
- Bacterial enteritis is a frequent cause of infectious diarrhea worldwide.
- Pathogenic enterobacteria, including Salmonella, Shigella, and E. coli O157:H7, are primary etiologic agents.
- Rapid and accurate diagnostic tools are crucial for effective treatment and control.
Purpose of the Study:
- To develop a rapid and definitive diagnostic test for bacterial enteritis.
- To detect and identify key pathogenic enterobacteria directly from clinical samples.
Main Methods:
- Conventional PCR assays were employed to detect Salmonella, Shigella, and E. coli O157:H7.
- Primers were designed targeting specific genes: uidA (general enterobacteria), ipaH (Shigella), 16SrRNA (Salmonella), and eaeA (E. coli O157:H7).
- The detection system incorporated common PCR, semi-nested PCR, and random amplified polymorphic DNA (RAPD).
Main Results:
- The PCR method demonstrated high sensitivity, specificity, and efficiency.
- Detection limits ranged from 3 to 50 CFU for the targeted pathogens.
- The entire detection process was completed within 4 hours.
Conclusions:
- The developed PCR method is a practical and routine protocol for pathogen detection.
- It enables rapid and accurate identification of bacterial enteritis-causing microorganisms from clinical samples.
- This advancement supports timely diagnosis and management of infectious enteritis.
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