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A Multiplex PCR Assay for the Rapid and Sensitive Detection of Methicillin-Resistant Staphylococcus aureus and Simultaneous Discrimination of Staphylococcus aureus from Coagulase-Negative Staphylococci

Authors

  • Benjin Xu,

    1. Author Xu is with College of Veterinary Medicine and College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Authors Ling Liu and Xiaofang Li are with College of Life Science and Chemistry, Tianshui Normal Univ., Tianshui, Gansu 741001, China. Authors Li Liu and Wang are with College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Xinping Li is with College of Veterinary Medicine, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Direct inquiries to author Xu (E-mail: xbj262020105@163.com).
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  • Ling Liu,

    1. Author Xu is with College of Veterinary Medicine and College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Authors Ling Liu and Xiaofang Li are with College of Life Science and Chemistry, Tianshui Normal Univ., Tianshui, Gansu 741001, China. Authors Li Liu and Wang are with College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Xinping Li is with College of Veterinary Medicine, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Direct inquiries to author Xu (E-mail: xbj262020105@163.com).
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  • Li Liu,

    1. Author Xu is with College of Veterinary Medicine and College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Authors Ling Liu and Xiaofang Li are with College of Life Science and Chemistry, Tianshui Normal Univ., Tianshui, Gansu 741001, China. Authors Li Liu and Wang are with College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Xinping Li is with College of Veterinary Medicine, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Direct inquiries to author Xu (E-mail: xbj262020105@163.com).
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  • Xinping Li,

    1. Author Xu is with College of Veterinary Medicine and College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Authors Ling Liu and Xiaofang Li are with College of Life Science and Chemistry, Tianshui Normal Univ., Tianshui, Gansu 741001, China. Authors Li Liu and Wang are with College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Xinping Li is with College of Veterinary Medicine, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Direct inquiries to author Xu (E-mail: xbj262020105@163.com).
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  • Xiaofang Li,

    1. Author Xu is with College of Veterinary Medicine and College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Authors Ling Liu and Xiaofang Li are with College of Life Science and Chemistry, Tianshui Normal Univ., Tianshui, Gansu 741001, China. Authors Li Liu and Wang are with College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Xinping Li is with College of Veterinary Medicine, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Direct inquiries to author Xu (E-mail: xbj262020105@163.com).
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  • Xin Wang

    1. Author Xu is with College of Veterinary Medicine and College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Authors Ling Liu and Xiaofang Li are with College of Life Science and Chemistry, Tianshui Normal Univ., Tianshui, Gansu 741001, China. Authors Li Liu and Wang are with College of Food Science and Engineering, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Xinping Li is with College of Veterinary Medicine, Northwest A & F Univ., Yangling, Shaanxi 712100, China. Direct inquiries to author Xu (E-mail: xbj262020105@163.com).
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Errata

This article is corrected by:

  1. Errata: Retraction Notice Volume 79, Issue 1, iv, Article first published online: 24 January 2014

Abstract

Abstract:  Methicillin-resistant Staphylococcus aureus (MRSA) is a global health concern, which had been detected in food and food production animals. Conventional testing for detection of MRSA takes 3 to 5 d to yield complete information of the organism and its antibiotic sensitivity pattern. So, a rapid method is needed to diagnose and treat the MRSA infections. The present study focused on the development of a multiplex PCR assay for the rapid and sensitive detection of MRSA. The assay simultaneously detected 4 genes, namely, 16S rRNA of the Staphylococcus genus, femA of S. aureus, mecA that encodes methicillin resistance, and one internal control. It was rapid and yielded results within 4 h. The analytical sensitivity and specificity of the multiplex PCR assay was evaluated by comparing it with the conventional method. The analytical sensitivity of the multiplex PCR assay at the DNA level was 10 ng DNA. The analytical specificity was evaluated with 10 reference staphylococci strains and was 100%. The diagnostic evaluation of MRSA was carried out using 360 foodborne staphylococci isolates, and showed 99.1% of specificity, 96.4% of sensitivity, 97.5% of positive predictive value, and 97.3% of negative predictive value compared to the conventional method. The inclusion of an internal control in the multiplex PCR assay is important to exclude false-negative cases. This test can be used as an effective diagnostic and surveillance tool to investigate the spread and emergence of MRSA.

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