A pilot study on PCR-based detection of four foodborne pathogenic microorganisms

  • Qiang GongEmail author
  • Zhanli Li
  • Mingfu Niu
Original Paper


To establish PCR-based detection methods for Staphylococcus aureus, Shigella, Pasteurella multocida and Pseudomonas aeruginosa, the nuc, ipah, ptfa and oprl genes were amplified by singleplex PCRs and multiplex PCR using specific primers that were designed according to the DNA sequences retrieved from GenBank. Then the annealing temperature was optimized, accompanied by a study of the specificity and sensitivity of the singleplex PCRs and multiplex PCR. The results showed that DNA fragments of 280, 474, 150 and 331 bp were specifically amplified from the four pathogenic bacteria mentioned above. No target DNA fragments were obtained from other pathogenic bacteria, including Salmonella typhimurium, Campylobacter jejuni, Clostridium perfringens and pathogenic Escherichia coli. The sensitivity of the singleplex PCRs were 100, 1, 1 and 10 pg/μL respectively. The detection limits of the four pathogenic bacteria in the multiplex PCR were 100, 1, 10 and 10 pg/µL respectively. These results showed that singleplex PCRs and multiplex PCR have good specificity and sensitivity. In conclusion, this experiment has laid a foundation for further research on rapid detection methods against these four pathogenic bacteria in food.


Foodborne pathogenic microorganism Polymerase chain reaction Singleplex Multiplex 



The authors gratefully acknowledge the grant from the production-study-research cooperation projects of Henan Province, China (No. 2014HNCXY008).


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© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  1. 1.College of Food and BioengineeringHenan University of Science and TechnologyLuoyangPeople’s Republic of China
  2. 2.Henan Engineering Laboratory of Livestock Disease Diagnosing and Food Safety TestingLuoyangPeople’s Republic of China
  3. 3.Henan Engineering Research Center of Food MaterialLuoyangPeople’s Republic of China

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