8株产NDM-1肠杆菌科细菌的耐药特点和流行性分析

doi:10.3969/j.issn.1673-8640.2018.07.009

摘要/Abstract

摘要：

目的研究温州医科大学附属第二医院临床分离的8株产新德里金属β-内酰胺酶1（NDM-1）肠杆菌科细菌的耐药特点及分子生物学特点。方法采用Vitek 2 Compact全自动微生物分析系统进行菌株的鉴定和体外药物敏感性试验;采用聚合酶链反应（PCR）扩增β-内酰胺酶基因,并对扩增产物进行测序和Blast比对分析。采用接合转移试验检测NDM-1耐药基因的水平转移能力。采用脉冲场凝胶电泳（PFGE）分析菌株的同源性。结果 8株菌株对β-内酰胺类药物及β-内酰胺酶抑制剂复合物均表现出较高的耐药性,而对氨基糖苷类和氟喹诺酮类药物敏感性很好;各菌株多同时携带多种耐药基因,其中5株携带SHV基因、2株携带CTX-M-1基因、1株携带DHA-1基因。产NDM-1菌株（2株大肠埃希菌除外）接合成功,接合子对β-内酰胺类药物的耐药性很高。PFGE结果显示,2株大肠埃希菌为不同克隆株,而3株阴沟肠杆菌中有2株具有同源性,视为同一克隆株。结论产NDM-1肠杆菌科细菌多同时携带多种耐药基因,对β-内酰胺类药物及β-内酰胺酶抑制剂复合物均表现出较高的耐药性。NDM-1耐药基因具有水平转移能力。

关键词: 新德里金属β-内酰胺酶1;, 碳青霉烯酶, 肠杆菌科, 脉冲场凝胶电泳

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

中图分类号:

R446.5

张盼, 沈振华, 张燕华, 刘兴晖, 李向阳. 8株产NDM-1肠杆菌科细菌的耐药特点和流行性分析[J]. 检验医学, 2018, 33(7): 616-621.

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.

图/表 5

参考文献 22

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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