Ann Clin Microbiol.  2019 Mar;22(1):9-13. 10.5145/ACM.2019.22.1.9.

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

• Kim SH
• Bae IK
• Kim NY
• Song SA
• Kim S
• Jeong J
• Shin JH

• KMID: 2441651
• DOI: http://doi.org/10.5145/ACM.2019.22.1.9

Abstract

BACKGROUND
The isolation of carbapenemase-producing Enterobacteriaceae (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.

Keyword

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

MeSH Terms

Acinetobacter baumannii
Disease Outbreaks
Enterobacteriaceae*
Enterococcus faecalis
Escherichia coli
Klebsiella pneumoniae
Mortality
Multiplex Polymerase Chain Reaction*
Polymerase Chain Reaction
Pseudomonas aeruginosa
Sensitivity and Specificity

Figure

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  Fig. 1 Multiplex PCR for the detection of seven carbapenemase-producing Enterobacteriaceae genes. Abbreviations: M, molecular weight marker; N, negative control. Lane: IMP (180 bp), VIM (306 bp), NDM (410 bp), OXA-23 (505 bp), OXA-48 (602 bp), KPC (747 bp), and GES (855 bp).

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