Ann Lab Med 2017; 37(5): 398-407  
Multidrug Resistance Mechanisms of Carbapenem Resistant Klebsiella pneumoniae Strains Isolated in Chongqing, China
Jinrong Yan, M.D.*, Shuli Pu, M.D.*, Xiaojiong Jia, M.D., Xiuyu Xu, M.D., Shuangshuang Yang, M.D., Jing Shi, M.D., Shan Sun, M.D., and Liping Zhang, M.D.
Department of Clinical Laboratory, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China
Correspondence to: Liping Zhang
Department of Clinical Laboratory, The First Affiliated Hospital of Chongqing Medical University, No.1 Youyi Road, Yuzhong District, Chongqing 400016, P. R. China
Tel: +86-023-89012756 Fax: +86-023-89012513 E-mail: 1309898173@qq.com
*These authors contributed equally to this work.
Received: November 24, 2016; Revised: February 10, 2017; Accepted: May 22, 2017; Published online: September 1, 2017.
© The Korean Society for Laboratory Medicine. All rights reserved.

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Abstract
Background: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is considered a serious global threat. However, little is known regarding the multidrug resistance (MDR) mechanisms of CRKP. This study investigated the phenotypes and MDR mechanisms of CRKP and identified their clonal characteristics.
Methods: PCR and sequencing were utilized to identify antibiotic resistance determinants. Integron gene cassette arrays were determined by restriction fragment length polymorphism (RFLP) analysis. Multi-locus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) were used for epidemiological analysis. Plasmids were typed by using a PCR-based replicon typing and analyzed by conjugation and transformation assays.
Results: Seventy-eight strains were identified as resistant to at least one carbapenem; these CRKP strains had a high prevalence rate (38.5%, 30/78) of carbapenemase producers. Additionally, most isolates harbored MDR genes, including Extended spectrum β-lactamases (ESBLs), AmpC, and quinolone and aminoglycoside resistance genes. Loss of porin genes was observed, and Class 1 integron was detected in 66.7% of the investigated isolates. PFGE and MLST results excluded the occurrence of clonal dissemination among these isolates.
Conclusions: A high prevalence of NDM-1 genes encoding carbapenem resistance determinants was demonstrated among the K. pneumoniae isolates. Importantly, this is the first report of blaNDM-1 carriage in a K. pneumoniae ST1383 clone in China and of a MDR CRKP isolate co-harboring blaNDM-1, blaKPC-2, blaCTX-M, blaSHV, acc(6’)-Ib, rmtB, qnrB, and acc(6’)-Ib-cr.
Keywords: Carbapenem-resistant, Klebsiella pneumoniae, Carbapenemases, Extended spectrum β-lactamases, Molecular epidemiology


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