Research on the drug resistance mechanism of carbapenem-resistant Escherichia coli

doi:10.3969/j.issn.1673-8640.2016.01.013

Abstract

Abstract:

Objective To investigate the drug resistance characteristics and mechanism of carbapenem-resistant Escherichia coli in Henan Provincial People's Hospital. Methods The identification and antimicrobial susceptibility in vitro of the isolates from qualified clinical specimens were analyzed by BD Phoenix 100 microorganism analysis system. The isolates of carbapenem-resistant Escherichia coli were screened according to the Clinical and Laboratory Standards Institute （CLSI） standard. Modified Hodges test and meropenem（MEM）-ethylene diamine tetraacetic acid （EDTA）-Na₂ double-disk synergy test were used for phenotypic screening of the isolates. By polymerase chain reaction（PCR）,the resistance genes,bla_KPC,bla_NDM,bla_VIM,bla_IMP and bla_OXA_-48,in the isolates were detected. The transferability of bla_KPC in the bla_KPC-positive isolates was studied by conjugation test. Results A total of 8 carbapenem-resistant Escherichia coli were screened from the 1 238 isolates. In the 8 isolates,7 isolates were resistant to imipenem （IPM）and MEM,but 1 isolate was only resistant to MEM. All the 8 isolates were positive in the modified Hodges test, and 6 isolates were positive in the MEM-EDTA double-disk synergy test. There were 2 isolates carrying bla_KPC_-2 and 3 isolates carrying bla_IMP_-4. Conjugation test indicated that bla_KPC_-2 could be transferred to Escherichia coli. Conclusions Carbapenem-resistant Escherichia coli carrying bla_IMP_-4 is firstly reported in the hospital. The resistance mechanism of carbapenem-resistant Escherichia coli in the hospital is mainly carrying bla_KPC_-2 and bla_IMP_-4,and the bla_KPC_-2 gene could be horizontally transmitted by plasmid.

Key words: Escherichia coli, Carbapenem, Drug resistance mechanism

CLC Number:

R378.2

YAN Wenjuan, LI Yi, WANG Shanmei, SUN Mingjie. Research on the drug resistance mechanism of carbapenem-resistant Escherichia coli[J]. Laboratory Medicine, 2016, 31(1): 56-60.

Figures/Tables 6

References 14

1	CHEN YT,LIN JC,FUNG CP,et al.KPC-2-encoding plasmids from Escherichia coli and Klebsiella pneumoniae in Taiwan[J]. J Antimicrob Chemother,2014,69(3): 628-631.
2	NORDMANN P,NAAS T,POIREL L. Global spread of carbapenemase-producing Enterobacteriaceae[J]. Emerg Infect Dis,2011,17(10): 1791-1798.
3	CHMELNITSKY I,SHKLYAR M,HERMESH O,et al. Unique genes identified in the epidemic extremely drug-resistant KPC-producing Klebsiella pneumoniae sequence type 258[J]. J Antimicrob Chemother,2013,68(1): 74-83.
4	彭乃杰,林哈妮,夏菲,等. 碳青霉稀类耐药肺炎克雷伯菌耐药机制和多位点序列分型研究[J].检验医学,2013,28(9):862-863.
5	蒯守刚,王卫萍,裴豪,等. 一株同时携带质粒介导KPC-2、DHA-1基因对碳青霉烯类药物耐药的产气肠杆菌[J].检验医学,2014,29(4):375-379.
6	许俊红,王中全. 质粒介导的KPC-2型碳青霉烯酶水平传播机制的探讨[J]. 医药论坛杂志,2013,34(1):87-88.
7	Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing;twenty-third informational supplement[S]. M100-S23,CLSI,2013.
8	PICÃO RC,ANDRADE SS,NICOLETTI AG,et al. Metallo-beta-lactamase detection:comparative evaluation of double-disk synergy versus combined disk tests for IMP-,GIM-,SIM-,SPM-,or VIM-producing isolates[J]. J Clin Microbiol,2008,46(6): 2028-2037.
9	DALLENNE C,DA COSTA A,DECRÉ D,et al. Development of a set of multiplex PCR assays for the detection of genes encoding important beta-lactamases in Enterobacteriaceae[J]. J Antimicrob Chemother,2010 ,65(3):490-495.
10	RAHMAN M,SHUKLA SK,PRASAD KN,et al.Prevalence and molecular characterisation of New Delhi metallo-β-lactamases NDM-1,NDM-5,NDM-6 and NDM-7 in multidrug-resistant Enterobacteriaceae from India[J]. Int J Antimicrob Agents,2014,44(1):30-37.
11	QUEENAN AM,BUSH K. Carbapenemases: the versatile beta-lactamases[J].Clin Microbiol Rev,2007,20(3):440-458.
12	MENA A,PLASENCIA V,GARCÍA L,et al. Characterization of a large outbreak by CTX-M-1-producing Klebsiella pneumoniae and mechanisms leading to in vivo carbapenem resistance development[J]. J Clin Microbiol,2006,44(8):2831-2837.
13	YANO H,KUGA A,OKAMOTO R,et al. Plasmid-encoded metallo-beta-lactamase(IMP-6) conferring resistance to carbapenems,especially meropenem[J].Antimicrob Agents Chemother,2001,45(5):1343-1348.
14	NORDMANN P,GNIADKOWSKI M,GISKE CG,et al. Identification and screening of carbapenemase-producing Enterobacteriaceae[J]. Clin Microbiol Infect,2012,18(5):432-438.

引物名称	序列（5'-3'）	片段（bp）	参考文献
KPCF	CATTCAAGGGCTTTCTTGCTGC	538	[9]
KPCR	ACGACGGCATAGTCATTTGC
NDMF	CACCTCATGTTTGAATTCGCC	984	[10]
NDMR	CTCTGTCACATCGAAATCGC
IMP-1F	TGAGCAAGTTATCTGTATTC	740	[11]
IMP-1R	TTAGTTGCTTGGTTTTGATG
VIM-1F	TTATGGAGCAGCAACCGATGT	920	[11]
VIM-1R	CAAAAGTCCCGCTCCAACGA
OXA-48F	TTGGTGGCATCGATTATCGG	744	[11]
OXA-48R	GAGCACTTCTTTTGTGATGGC

引物名称	序列（5'-3'）	片段（bp）	参考文献
KPCF	CATTCAAGGGCTTTCTTGCTGC	538	[9]
KPCR	ACGACGGCATAGTCATTTGC
NDMF	CACCTCATGTTTGAATTCGCC	984	[10]
NDMR	CTCTGTCACATCGAAATCGC
IMP-1F	TGAGCAAGTTATCTGTATTC	740	[11]
IMP-1R	TTAGTTGCTTGGTTTTGATG
VIM-1F	TTATGGAGCAGCAACCGATGT	920	[11]
VIM-1R	CAAAAGTCCCGCTCCAACGA
OXA-48F	TTGGTGGCATCGATTATCGG	744	[11]
OXA-48R	GAGCACTTCTTTTGTGATGGC

菌株	科室	年龄（岁）	性别	样本	IPM （μg/mL）	MEM （μg/mL）	Hodges 试验	双纸片协同试验	bla_KPC_-2 基因	bla_IMP_-4 基因
ECO1	ICU	80	男	血液	>32	>32	+	–	+	–
ECO2	神经内科	83	男	痰液	>32	>32	+	–	+	–
ECO3	儿科	9个月	男	尿液	>32	>32	+	+	–	–
ECO4	儿科	9个月	男	血液	>32	>32	+	+	–	–
ECO5	ICU	64	女	痰液	8	>32	+	+	–	+
ECO6	ICU	66	男	痰液	>32	>32	+	+	–	+
ECO7	呼吸科	70	女	痰液	>32	>32	+	+	–	–
ECO8	CCU	50	女	痰液	2	>32	+	+	–	+

菌株	科室	年龄（岁）	性别	样本	IPM （μg/mL）	MEM （μg/mL）	Hodges 试验	双纸片协同试验	bla_KPC_-2 基因	bla_IMP_-4 基因
ECO1	ICU	80	男	血液	>32	>32	+	–	+	–
ECO2	神经内科	83	男	痰液	>32	>32	+	–	+	–
ECO3	儿科	9个月	男	尿液	>32	>32	+	+	–	–
ECO4	儿科	9个月	男	血液	>32	>32	+	+	–	–
ECO5	ICU	64	女	痰液	8	>32	+	+	–	+
ECO6	ICU	66	男	痰液	>32	>32	+	+	–	+
ECO7	呼吸科	70	女	痰液	>32	>32	+	+	–	–
ECO8	CCU	50	女	痰液	2	>32	+	+	–	+

[1]	WANG Jiawei, ZHU Weinan, CHEN Yingying, JI Ping, WANG Ying. Clinical characteristics，molecular typing and drug resistance genes of carbapenem-resistant Klebsiella pneumoniae isolated from blood culturing [J]. Laboratory Medicine, 2023, 38(4): 362-367.
[2]	XU Qi, PAN Fen, SUN Yan, SHI Yingying, YU Fangyuan, ZHANG Hong. Methodology evaluation of screening techniques for CRE colonized in intestinal tract [J]. Laboratory Medicine, 2022, 37(8): 761-765.
[3]	WEN Qinghui, LI Fengying, LI Yanchang, HUANG Jinyin, WANG Fengping. Drug susceptibility of multidrug-resistant ESBL-producing Escherichia coli to antimicrobial agents restored by different concentrations of EDTA [J]. Laboratory Medicine, 2022, 37(3): 205-208.
[4]	PAN Yunqi, LI Yungai, WANG Jianqiang, WU Qiong, TANG Jin. Analysis of molecular typing,virulence genes and antibiotic susceptibility of uropathogenic Escherichia coli in patients with recurrent urinary tract infection [J]. Laboratory Medicine, 2022, 37(2): 141-145.
[5]	SUN Ying, LI Yi, YAN Wenjuan, JING Nan, MA Bing. Analysis of drug resistance of rare CRE and clinical application of carbapenem screening test in Henan [J]. Laboratory Medicine, 2022, 37(2): 146-149.
[6]	BAO Hailin, HUA Hongyan, SUN Hengliang, LIU Hua, JI Shunnian, QIN Chenhao, DU Hong. Rapid identification of carbapenemase producing Enterobacteriaceae phenotypes by modified Carba Np test and mCIM/eCIM test [J]. Laboratory Medicine, 2022, 37(10): 963-968.
[7]	WANG Lixin, SU Yunfu, WANG Yi. Research progress on the drug resistance mechanism of clinical pathogens to fosfomycin [J]. Laboratory Medicine, 2022, 37(10): 993-997.
[8]	YAO Lifeng, ZHENG Qiaoping, YANG Meng, LUO Qingqiong, CHEN Xu, CHEN Fuxiang. Evaluation of colloidal gold immunochromatography assay for the rapid determination of carbapenemase in CRE isolates [J]. Laboratory Medicine, 2022, 37(1): 68-71.
[9]	YIN Juan, WANG Yingchao, SUI Yang, JIANG Chun, SHI Yunqi, ZHU Chaowang. Efficacy evaluation and application value of screening carbapenemase in enterobacteriaceae by mCIM and eCIM [J]. Laboratory Medicine, 2021, 36(2): 177-180.
[10]	YANG Hongmei, TAO Jianmin, WEI Tian, WANG Jiangjun, MAO Jiahong, WANG Haiying. Incidenceand resistance profile of carbapenem-resistant Enterobacteriaceae among inpatients in a clinical center in Shanghai from 2015 to 2019 [J]. Laboratory Medicine, 2021, 36(1): 87-91.
[11]	LU Shuhua, LI Xiaozhe, LIU Lingyun, JIN Chengqiang, GE Ningning, DONG Haixin. Clinical infection characteristics and their drug resistance genes of carbapenem-resistant Enterobacteriaceae in a hospital of Shandong [J]. Laboratory Medicine, 2020, 35(8): 757-762.
[12]	LIU Yan, LONG Yin, PAN Weijie, ZHANG Tong, WENG Wenhao, YU Ying. Role of fecal DNA level of Fusobacterium nucleatum and clbA+ Escherichia coli in the diagnosis of colorectal cancer [J]. Laboratory Medicine, 2020, 35(4): 314-317.
[13]	WANG Junjie, YAO Zonghui, MA Bing, LI Yi, YAN Wenjuan, WANG Shanmei, MA Qiong, YUAN Youhua. Risk factors and prognosis of bloodstream infection caused by carbapenem-resistant Klebsiella pneumoniae [J]. Laboratory Medicine, 2020, 35(3): 195-199.
[14]	GAO Juyi, WU Chuanan, YANG Weikang, LUO Yuxuan, LIU Shengnan, XU Xiaoping. Role of COP microfluidic chip for the rapid detection of Escherichia coli O157:H7 [J]. Laboratory Medicine, 2020, 35(2): 148-152.
[15]	LIU Cunfen, WANG Yan, YANG Rong, HU Lixiang. Relationship between ESBL genotyping of Escherichia coli and integron in Anning [J]. Laboratory Medicine, 2020, 35(12): 1263-1266.