A clinical isolate of carbapenem-resistant Enterobacter aerogen with plasmid-mediated KPC-2 and DHA-1 genes

doi:10.3969/j.issn.1673-8640.2014.04.017

Abstract

Abstract:

Objective　To investigate the resistance and transmission mechanisms of a clinical isolate of carbapenem-resistant Enterobacter aerogen. Methods　The minimal inhibition concentrations（MIC） of antimicrobial agents were determined by agar dilution method， and plasmid conjugation experiment， plasmid extraction and DNA molecular hybridization， isoelectric focusing electrophoresis（IEF）， polymerase chain reaction（PCR）， DNA sequencing and outer-membrane protein analysis were used for analyzing the resistant gene and transmission mechanism. Results IEF showed that there were 3 bands of beta-lactamases with isoelectric point（pI） of 5.4， 6.7 and 7.8 in the clinical isolate of carbapenem-resistant Enterobacter aerogen. These 3 bands of beta-lactamases were confirmed to be TEM-1（pI 5.4）， KPC-2（pI 6.7） and DHA-1（pI 7.8） by PCR amplification and DNA sequencing， and 2 of them（pIs of 6.7 and 7.8） were found that they can be transferred by plasmid conjugantion experiment. KPC-2 and DHA-1 genes were located on an about 56 kb plasmid by plasmid conjugation experiment， plasmid extraction and DNA molecular hybridization. Outer-membrane protein electrophoresis analysis revealed that a 41 000 outer-membrane protein was absent in the clinical isolate of carbapenem-resistant Enterobacter aerogen comparing with clinical wild-type Enterobacter aerogen. Conclusions The clinical isolate of Enterobacter aerogen resistant to carbapenem produces a plasmid-mediaed carbapemase KPC-2 which belongs to Group 2f， Class A beta-lactamase. DHA-1 enzyme with outer-membrane protein absence may be related with the resistant mechanism of carbapenem-resistance in the isolate of Enterobacter aerogen.

Key words: Enterobacter aerogen, Carbapenem, Plasmid, Beta-lactamase, Enterobacteriaceae

CLC Number:

R378.2

KUAI Shougang，WANG Weiping，PEI Hao，FAN Ming，LIU Jun，ZHOU Xike，SHANG Zhongbo，SHAO Haifeng. A clinical isolate of carbapenem-resistant Enterobacter aerogen with plasmid-mediated KPC-2 and DHA-1 genes[J]. , 2014, 29(4): 375-379.

References

[1] Queenan AM,Bush K. Carbapenemases: the versatile beta-lactamases[J]. Clin Microbiol Rev, 2007, 20(3): 440-458.
[2] Miriagou V,Cornaglia G, Edelstein M, et al. Acquired carbapenemases in Gram-negative bacterial pathogens: detection and surveillance issues[J]. Clin Microbiol Infect, 2010, 16(2):112-122.
[3] Nordmann P,Cuzon G, Naas T. The real threat of Klebsiella pneumoniae carbapenemase-producing bacteria[J]. Lancet Infect Dis, 2009, 9(4):228-236.
[4] Hanson ND , Moland ES, Hossain A, et al. Unusual Salmonella enterica serotype Typhimurium isolate producing CMY-7, SHV-9 and OXA-30 beta-lactamases[J]. J Antimicrob Chemother, 2002, 49(6):1011-1014.
[5] Héritier C, Poirel L, Aubert D, et al. Genetic and functional analysis of the chromosome-encoded carbapenem-hydrolyzing oxacillinase OXA-40 of Acinetobacter baumannii[J] . Antimicrob Agents Chemother, 2003,47(1): 268-273.
[6] 蒯守刚,邵海枫,王卫萍,等.耐碳青酶烯类肠杆菌科细菌的耐药机制研究[J]. 中华医院感染学杂志, 2010, 20(22):3441-3444.
[7] Smith Moland E, Hanson ND, Herrera VL, et al. Plasmid-mediated carbapenem-hydrolysing beta-lactamase, KPC-2, in Klebsiella pneumoniae isolates[J]. Antimicrob Agents Chemother, 2003, 51(3): 711-714.
[8] Nüesch-Inderbinen MT, Hchler H, Kayser FH. Detection of genes coding extended-spectrum SHV beta-lactamases in clinical isolates by a molecular genetic method and comparison with the E test[J]. Eur J Clin Microbiol Infect Dis, 1996, 15(5): 398-402.
[9] Rasheed JK, Jay C, Metchock B, et al. Evolution of extended-spectrum beta-lactam resistance (SHV-8) in a strain of Escherichia coli during multiple episodes of bacteremia[J]. Antimicrob Agents Chemother, 1997, 41(3): 647-653.
[10] Edelstein M, Pimkin M,Palagin I,et al. Prevalence and molecular epidemiology of CTX-M extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in Russian hospitals[J]. Antimicrob Agents Chemother, 2003, 47(12): 3724-3732.
[11] Grimont F, Grimont PAD. Prokaryotes: a handbook on the biology of bacteria[M]. 3rd ed. New York: Springer, 2006: 219-244.
[12] 蒯守刚,裴豪,陈燕燕,等.亚胺培南耐药大肠埃希菌基因型检测[J].检验医学, 2011,26(10): 653-657.
[13] 蒯守刚,邵海枫,王卫萍,等.大肠埃希菌对碳青霉烯类药物的耐药机制研究[J].中华微生物学和免疫学杂志,2010,30(9):829-833.
[14] Chenia HY, Pillay B, Pillay D. Analysis of the mechanisms of fluoroquinolone resistance in urinary tract pathogens[J]. J Antimicrob Chemother,2006,58(6):1274-1278.

[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]	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.
[5]	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.
[6]	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.
[7]	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.
[8]	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.
[9]	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.
[10]	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.
[11]	WANG Su, ZHAO Hu. Development and application of clinical determination and treatment for carbapenem-resistant Enterobacteriaceae [J]. Laboratory Medicine, 2020, 35(10): 967-970.
[12]	ZHOU Hongwei, HU Yanyan, ZHANG Rong. Determination methods for carbapenem-resistant Enterobacteriaceae [J]. Laboratory Medicine, 2020, 35(10): 971-973.
[13]	LI Shirong, LIN Lijing, JIANG Xiaofei, GUAN Ming. Role of double concentration combined modified carbapenem inactivation test for screening CRE [J]. Laboratory Medicine, 2020, 35(10): 979-982.
[14]	HE Lihua, NI Lijun, YANG Simin, YU Xiaoyu, ZHOU Aiping, HU Liang, GUO Jian, WU Wenjuan. Clinical application of RT-PCR for Klebsiella pneumoniae producing KPC carbapenemase [J]. Laboratory Medicine, 2020, 35(10): 983-987.
[15]	CHEN Tao, XU Ye, DONG Guofeng, ZHOU Cui, XU Wenya, ZHOU Tieli. In vitro antibacterial activity of ceftazidime/avibactam alone and in combination with fosfomycin against carbapenem-resistant Gram-negative bacilli [J]. Laboratory Medicine, 2020, 35(10): 988-993.