Drug resistance characteristics and epidemiological analysis of 8 isolates of NDM-1-producing Enterobacteriaceae

doi:10.3969/j.issn.1673-8640.2018.07.009

Abstract

Abstract:

Objective To investigate the drug resistance and epidemiological characteristics of 8 isolates of New Delhi metallo-β-lactamase 1（NDM-1）-producing Enterobacteriaceae isolated from the Second Affiliated Hospital of Wenzhou Medical University. Methods Vitek 2 Compact automatic microbiological system was used for bacterial identification and antimicrobial susceptibility test. β-lactamase genes were determined by polymerase chain reaction （PCR）,which were performed sequencing and BLAST comparison analysis. The transferability of NDM-1 gene was determined by conjugational transfer experiment. The homology was analyzed by pulsed-field gel electrophoresis （PFGE）. Results The 8 isolates showed high drug resistance to β-lactam antibiotics and β-lactamase inhibitor combinations,and they had relatively better susceptibility to aminoglycosides and fluoroquinolones. Most isolates carried multiple drug resistance genes,including 5 isolates harboured SHV gene,2 isolates harboured CTX-M-1 gene,and 1 isolate harboured DHA-1 gene. Transconjugants were successfully obtained from all NDM-1-producing isolates except 2 isolates of Escherichia coli. All transconjugants showed high drug resistance to β-lactam antibiotics. PFGE showed that 2 isolates of Escherichia coli belonged to different clone isolates,and 2 from 3 isolates of Enterobacter cloacae had homology,which was seen as the same clone. Conclusions Most NDM-1-producing isolates carry multiple drug resistance genes at the same time,which show high drug resistance to β-lactam antibiotics and β-lactamase inhibitor combinations. NDM-1 gene has horizontal transferability in different isolates.

Key words: New Delhi metallo-β-lactamase 1;, Carbapenemase, Enterobacteriaceae, Pulsed-field gel electrophoresis

CLC Number:

R446.5

ZHANG Pan, SHEN Zhenhua, ZHANG Yanhua, LIU Xinghui, LI Xiangyang. Drug resistance characteristics and epidemiological analysis of 8 isolates of NDM-1-producing Enterobacteriaceae[J]. Laboratory Medicine, 2018, 33(7): 616-621.

Figures/Tables 5

References 22

[1]	YONG D, TOLEMAN M A, GISKE C G, et al.Characterization of a new metallo-beta-lactamase gene, bla(NDM-1), and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India[J]. Antimicrob Agents Chemother, 2009, 53(12): 5046-5054.
[2]	郝琼,刘翔,郭邦成,等. 宁夏腹泻病人群携带NDM-1基因菌株的初步研究[J]. 宁夏医学杂志, 2012, 34(4) : 289-291.
[3]	Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing[S]. M100-S20,CLSI,2013.
[4]	NORDMANN P, POIREL L, CARRËR A, et al. How to detect NDM-1 producers[J]. J Clin Microbiol, 2011, 49(2):718-721.
[5]	KUMARASAMY K K, TOLEMAN M A, WALSH T R, et al.Emergence of a new antibiotic resistance mechanism in India, Pakistan, and the UK: a molecular, biological, and epidemiological study[J]. Lancet Infect Dis, 2010, 10(9): 597-602.
[6]	POTRON A, POIREL L,NORDMANN P.Plasmid-mediated transfer of the bla(NDM-1) gene in Gram-negative rods[J]. FEMS Microbiol Lett, 2011, 324(2):111-116.
[7]	陈硕,邱少富,夏力亮,等. 鲍曼不动杆菌blaNDM-1基因序列分析及表达[J]. 中国人兽共患病学报, 2012, 28(5): 471-473.
[8]	刘洋,李方去,蒋伟燕,等. 质粒介导KPC-2型碳青霉烯酶肺炎克雷伯菌儿童分离株耐药基因研究[J]. 中华微生物学和免疫学杂志,2012,32(10):861-865.
[9]	WANG J, YUAN M, CHEN H, et al. First report of Klebsiella oxytoca strain simultaneously producing NDM-1, IMP-4, KPC-2 carbapenemases[J]. Antimicrob Agents Chemother, 2017, 61(9): pii:e00877-17.
[10]	ZHENG R, ZHANG Q, GUO Y, et al.Outbreak of plasmid-mediated NDM-1-producing Klebsiella pneumoniae ST105 among neonatal patients in Yunnan, China[J]. Ann Clin Microbiol Antimicrob, 2016, 15:10.
[11]	LIU Y, WAN L G, DENG Q, et al.First description of NDM-1-, KPC-2-, VIM-2- and IMP-4- producing Klebsiella pneumoniae strains in a single Chinese teaching hospital[J]. Epidemiol Infect, 2015, 143(2):376-384.
[12]	曹丽军,李慧卿,耿凤珍,等. 两株携带NDM-1型金属β-内酰胺酶阴沟肠杆菌的临床特征研究[J]. 中华医院感染学杂志, 2015, 25(23): 5305-5307.
[13]	朱水荣,商小春,帅慧群,等. 浙江省首次检出1株携带NDM-1基因的肺炎克雷伯菌[J]. 中国人兽共患病学报, 2015, 31(1):30-34.
[14]	CHEN Y, ZHOU Z, JIANG Y, et al.Emergence of NDM-1-producing Acinetobacter baumannii in China[J]. J Antimicrob Chemother, 2011, 66(6):1255-1259.
[15]	HO P L, LO W U, YEUNG M K, et al.Complete sequencing of pNDM-HK encoding NDM-1 carbapenemase from a multidrug-resistant Escherichia coli strain isolated in Hong Kong[J]. PLoS One, 2011, 6(3): e17989.
[16]	VILLA L, GUERRA B, SCHMOGER S, et al.IncA/C plasmid carrying bla(NDM-1), bla(CMY-16), and fosA3 in a Salmonella enterica serovar corvallis strain isolated from a migratory wild bird in germany[J]. Antimicrob Agents Chemother, 2015, 59(10): 6597-6600.
[17]	POIREL L, BONNIN R A, NORDMANN P.Analysis of the resistome of a multidrug-resistant NDM-1-producing Escherichia coli strain by high-throughput genome sequencing[J]. Antimicrob Agents Chemother, 2011, 55(9):4224-4229.
[18]	MATASEJE L F, BOYD D A, LEFEBVRE B, et al.Complete sequences of a novel blaNDM-1- harbouring plasmid from Providencia rettgeri and an FII-type plasmid from Klebsiella pneumoniae identified in Canada[J]. J Antimicrob Chemother, 2014, 69(3):637-642.
[19]	VILLA L, POIREL L, NORDMANN P, et al.Complete sequencing of an IncH plasmid carrying the blaNDM-1, blaCTX-M-15 and qnrB1 genes[J]. J Antimicrob Chemother, 2012, 67(7):1645-1650.
[20]	DOLEJSKA M, VILLA L, POIREL L, et al.Complete sequencing of an IncHI1 plasmid encoding the carbapenemase NDM-1, the ArmA 16S RNA methylase and a resistance-nodulation-cell division/multidrug efflux pump[J]. J Antimicrob Chemother, 2013, 68(1):34-39.
[21]	HU H, HU Y, PAN Y, et al.Novel plasmid and its variant harboring both a bla(NDM-1) gene and type Ⅳ secretion system in clinical isolates of Acinetobacter lwoffii[J]. Antimicrob Agents Chemother, 2012, 56(4): 1698-1702.
[22]	GUO Q, SU J, MCELHENY C L, et al. IncX2 and IncX1-X2 hybrid plasmids coexisting in a FosA6-producing Escherichia coli strain[J]. Antimicrob Agents Chemother, 2017, 61(7): pii:e00536-17.

菌株编号	样本来源	感染高危因素						抗菌治疗	转归
菌株编号	样本来源	年龄	基础疾病	外科手术	气管插管	糖皮质激素	抗菌药物预防	抗菌治疗	转归
1	痰液	34岁	高血压、脑出血	√	√	√	√	依替米星、阿莫西林-克拉维酸	转院
2	血液	4个月	胆管炎、支气管炎	√	×	×	√	头孢曲松、甲硝唑	出院
3	痰液	2个岁	急性胃肠炎	×	√	×	√	左氧氟沙星、头孢噻肟	转院
4	痰液	10个月	惊厥	×	√	√	√	左氧氟沙星、头孢哌酮-舒巴坦	治愈
5	痰液	1岁	肺炎	√	√	×	√	左氧氟沙星	治愈
6	脓液	46岁	直肠癌、造瘘	√	√	×	√	左氧氟沙星	控制
7	尿液	4岁	重复肾、尿路感染	√	×	×	×	头孢哌酮-舒巴坦	治愈
8	尿液	63岁	高血压、糖尿病	×	×	×	×	左氧氟沙星	控制

菌株编号	样本来源	感染高危因素						抗菌治疗	转归
菌株编号	样本来源	年龄	基础疾病	外科手术	气管插管	糖皮质激素	抗菌药物预防	抗菌治疗	转归
1	痰液	34岁	高血压、脑出血	√	√	√	√	依替米星、阿莫西林-克拉维酸	转院
2	血液	4个月	胆管炎、支气管炎	√	×	×	√	头孢曲松、甲硝唑	出院
3	痰液	2个岁	急性胃肠炎	×	√	×	√	左氧氟沙星、头孢噻肟	转院
4	痰液	10个月	惊厥	×	√	√	√	左氧氟沙星、头孢哌酮-舒巴坦	治愈
5	痰液	1岁	肺炎	√	√	×	√	左氧氟沙星	治愈
6	脓液	46岁	直肠癌、造瘘	√	√	×	√	左氧氟沙星	控制
7	尿液	4岁	重复肾、尿路感染	√	×	×	×	头孢哌酮-舒巴坦	治愈
8	尿液	63岁	高血压、糖尿病	×	×	×	×	左氧氟沙星	控制

抗菌药物	MIC（μg/mL）
抗菌药物	1	2	3	4	5	6	7	8
阿米卡星	≤2S	≤2S	≤2S	≤2S	≤2S	≤2S	8S	≤2S
头孢唑啉	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R
头孢他啶	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R
头孢曲松	≥64R	≥64R	≥64R	≥64R	≥64R	16R	≥64R	≥64R
头孢吡肟	32R	≥64R	≥64R	≥64R	32R	2S	≤1S	≥64R
庆大霉素	≤1S	≤1S	≤1S	≤1S	≤1S	≤1S	≤1S	≤1S
氨曲南	≤1S	≥64R	≥64R	≥64R	16R	≤1S	16I	≥64R
环丙沙星	≥4R	≥4R	≤0.25S	≤0.25S	0.5S	1S	≥4R	0.5S
亚胺培南	4R	32R	32R	32R	16R	8R	4R	8R
左氧氟沙星	≥8R	≥8R	1S	1S	1S	1S	≥8R	1S
复方磺胺甲噁唑	≥320R	40S	≥320R	≥320R	≥320R	≤20S	≥320R	≥320R
氨苄西林-舒巴坦	≥32R	≥32R	≥32R	≥32R	≥32R	≥32R	≥32R	≥32R
哌拉西林-舒巴坦	64I	≥128R	≥128R	≥128R	≥128R	64I	32I	≥128R
妥布霉素	8I	≤1S	≤1S	≤1S	8I	8I	8I	≤1S
厄他培南	16R	32R	16R	8R	8R	4R	2R	16R
呋喃妥因	≥512R	256R	256R	128R	64I	≤16S	128R	≤16S
头孢替坦	≥64R	≥64R	≥64R	≥64R	≥64R	8S	≥64R	≥64R

抗菌药物	MIC（μg/mL）
抗菌药物	1	2	3	4	5	6	7	8
阿米卡星	≤2S	≤2S	≤2S	≤2S	≤2S	≤2S	8S	≤2S
头孢唑啉	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R
头孢他啶	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R	≥64R
头孢曲松	≥64R	≥64R	≥64R	≥64R	≥64R	16R	≥64R	≥64R
头孢吡肟	32R	≥64R	≥64R	≥64R	32R	2S	≤1S	≥64R
庆大霉素	≤1S	≤1S	≤1S	≤1S	≤1S	≤1S	≤1S	≤1S
氨曲南	≤1S	≥64R	≥64R	≥64R	16R	≤1S	16I	≥64R
环丙沙星	≥4R	≥4R	≤0.25S	≤0.25S	0.5S	1S	≥4R	0.5S
亚胺培南	4R	32R	32R	32R	16R	8R	4R	8R
左氧氟沙星	≥8R	≥8R	1S	1S	1S	1S	≥8R	1S
复方磺胺甲噁唑	≥320R	40S	≥320R	≥320R	≥320R	≤20S	≥320R	≥320R
氨苄西林-舒巴坦	≥32R	≥32R	≥32R	≥32R	≥32R	≥32R	≥32R	≥32R
哌拉西林-舒巴坦	64I	≥128R	≥128R	≥128R	≥128R	64I	32I	≥128R
妥布霉素	8I	≤1S	≤1S	≤1S	8I	8I	8I	≤1S
厄他培南	16R	32R	16R	8R	8R	4R	2R	16R
呋喃妥因	≥512R	256R	256R	128R	64I	≤16S	128R	≤16S
头孢替坦	≥64R	≥64R	≥64R	≥64R	≥64R	8S	≥64R	≥64R