Simultaneous detection of Escherichia coli O157:H7, Staphylococcus aureus and Salmonella by multiplex PCR in milk

Wei, Caijiao; Zhong, Junliang; Hu, Ting; Zhao, Xihong

doi:10.1007/s13205-018-1086-5

Original Article
Published: 13 January 2018

Simultaneous detection of Escherichia coli O157:H7, Staphylococcus aureus and Salmonella by multiplex PCR in milk

3 Biotech volume 8, Article number: 76 (2018) Cite this article

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Abstract

Escherichia coli O157:H7, Staphylococcus aureus, and Salmonella are food-borne pathogens that cause serious gastrointestinal illness and frequent food safety accidents. This study aimed to develop a practical multiplex polymerase chain reaction (mPCR) technique for the simultaneous detection of these food-borne pathogens in culture broth and artificial food matrix. Pathogen-specific DNA sequences in the rfbE, nuc, and invA genes were used as targets to design primers for the identification of E. coli O157:H7, S. aureus, and Salmonella, respectively. As expected, the method produced species-specific bands of amplified products without any contaminating non-specific bands. The highest species specificity was established with primer concentrations of 0.1, 0.2, and 0.4 μM for E. coli O157:H7, S. aureus, and Salmonella, correspondingly. The detection sensitivity of this assay was 10³ CFU/mL in culture broth, and the limit of detection was consistent with singleplex PCR in the food sample. The mPCR assay proposed here is an easy and convenient detection method, which will be valuable for microbial epidemiology and food safety investigations.

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Acknowledgements

This work has been supported by the National Natural Science Foundation of China (Grant no. 31501582) and open project program of key laboratory for Green Chemical Process of Ministry of Education in Wuhan Institute of Technology (2017007).

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Key Laboratory for Green Chemical Process of Ministry of Education, Key Laboratory for Hubei Novel Reactor & Green Chemical Technology, Research Center for Environmental Ecology and Engineering, School of Environmental Ecology and Biological Engineering, Wuhan Institute of Technology, Wuhan, 430205, China
Caijiao Wei, Junliang Zhong, Ting Hu & Xihong Zhao

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Caijiao Wei
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Junliang Zhong
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Ting Hu
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Xihong Zhao
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Correspondence to Xihong Zhao.

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Wei, C., Zhong, J., Hu, T. et al. Simultaneous detection of Escherichia coli O157:H7, Staphylococcus aureus and Salmonella by multiplex PCR in milk. 3 Biotech 8, 76 (2018). https://doi.org/10.1007/s13205-018-1086-5

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Received: 19 May 2017
Accepted: 02 January 2018
Published: 13 January 2018
DOI: https://doi.org/10.1007/s13205-018-1086-5

Keywords

Multiplex PCR
Escherichia coli O157:H7
Staphylococcus aureus
Salmonella
Detection method