Ann Lab Med 2019; 39(1): 36-42  https://doi.org/10.3343/alm.2019.39.1.36
Molecular Epidemiology and Mechanisms of 43 Low-Level Linezolid-Resistant Enterococcus faecalis Strains in Chongqing, China
Ruoyi Hua, M.D., Yun Xia, Ph.D., Wenyao Wu, M.D., Mi Yang, M.D., and Jia Yan, M.D.
Department of Clinical Laboratory, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China
Corresponding author: Yun Xia, Ph.D.
https://orcid.org/0000-0003-3136-8548
Department of Clinical Laboratory, The First Affiliated Hospital of Chongqing Medical University, 1 You Yi Road, Yuzhong District, Chongqing 40016, China
Tel: +86-023-89012742
Fax: +86-023-89012513
E-mail: xiayun12cn@aliyun.com
Received: December 6, 2017; Revised: April 12, 2018; Accepted: August 16, 2018; Published online: January 1, 2019.
© Korean Society for Laboratory Medicine. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background: Enterococcus faecalis strains with low-level resistance to linezolid (an oxazolidinone antibiotic) have become common. No large-scale study has examined the underlying mechanisms in linezolid-resistant E. faecalis (LRE) strains. We investigated these mechanisms and molecular characteristics in Chongqing, China.
Methods: A total of 1,120 non-duplicated E. faecalis strains collected from August 2014 to June 2017 underwent drug susceptibility testing. LRE strains were screened for optrA, cfr, and mutations in the 23S rRNA and ribosomal proteins L3 and L4 by PCR amplification and sequencing. Multi-locus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) were used for epidemiological analysis.
Results: All 43 low-level LRE strains (minimum inhibitory concentration: 8–16 mg/L) harbored optrA; cfr and 23S rRNA mutations were not detected. Novel mutations in the ribosomal proteins L3 and L4-one deletion (Q103del) and four substitutions (S113L, T35A, I98V, and N79D)-were identified. Novel amino acid substitutions at positions E60K, G197D, and T285P of the OptrA protein were observed. MLST revealed 20 types of LRE strains; the most common type was ST16 (32.6%). PFGE showed 14 strains of ST16 with unique banding patterns. Eight novel sequence types (ST823 to ST830) and one allele (gki95) were identified for the first time in China.
Conclusions: optrA plays an important role in linezolid resistance and may serve as a marker for resistance screening. Since the L3 and L4 mutations did not simultaneously occur in the same strain, they play a negligible role in linezolid resistance. Epidemiological investigation suggested that the LRE cases were sporadic.
Keywords: Linezolid, Resistance, Mechanism, Enterococcus, Epidemiology



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