Annals of Clinical Microbiology, The official Journal of the Korean Society of Clinical Microbiology

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pISSN 2288-0585 eISSN 2288-6850

Development and Evaluation of Multiplex PCR for the Detection of Carbapenemase-Producing Enterobacteriaceae

Original article

Annals of Clinical Microbiology (Ann Clin Microbiol) 2019 March, Volume 22, Issue 1, Pages 9-13.

https://doi.org/10.5145/ACM.2019.22.1.9

Development and Evaluation of Multiplex PCR for the Detection of Carbapenemase-Producing Enterobacteriaceae

Si Hyun Kim1*, Il Kwon Bae2*, Na Young Kim3,4, Sae Am Song3, Sunjoo Kim5, Joseph Jeong6, Jeong Hwan Shin3,4

1Department of Clinical Laboratory Science, Semyung University, Jecheon, 2Department of Dental Hygiene, Silla University, 3Department of Laboratory Medicine, Inje University College of Medicine, 4Paik Institute for Clinical Research, Inje University College of Medicine, Busan, 5Department of Laboratory Medicine, Gyeongsang National University College of Medicine, Jinju, 6Department of Laboratory Medicine, Ulsan University Hospital, University of Ulsan College of Medicine, Ulsan, Korea

Abstract

Background: The isolation of carbapenemase-producing  (CPE) has become increasingly common. Continuous surveillance for these organisms is essential because their infections are closely related to outbreaks of illness and are associated with high mortality rates. The aim of this study was to develop and evaluate multiplex PCR as a means of detecting several important CPE genes simultaneously.

Methods: We aimed to develop a multiplex PCR that could detect seven CPE genes simultaneously. The multiplex PCR was composed of seven primer sets for the detection of KPC, IMP, VIM, NDM-1, GES, OXA-23, and OXA-48. We designed different PCR product sizes of at least 100 bp. We evaluated the performance of this new test using 69 CPE-positive clinical isolates. Also, we confirmed the specificity to rule out false-positive reactions by using 71 carbapenem-susceptible clinical strains.

Results: A total of 69 CPE clinical isolates showed positive results and were correctly identified as KPC (N=14), IMP (N=13), OXA-23 (N=12), OXA-48 (N=11), VIM (N=9), GES (N=5), and NDM (N=5) by the multiplex PCR. All 71 carbapenem-susceptible clinical isolates, including Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa, showed negative results.

Conclusion: This multiplex PCR can detect seven CPE genes at a time and will be useful in clinical laboratories. (Ann Clin Microbiol 2019;22:9-13)

Keywords

Carbapenem, Carbapenem-resistant Enterobacteriaceae, Carbapenemase-producing Enterobacteriaceae, Enterobacteriaceae, Multiplex PCR