临床分离万古霉素耐药肠球菌van基因型及分子分型

中国感染与化疗杂志2018年1月20日第18卷第1期?Chin?J?Infect?Chemother,?January.?2018,?Vol.?18,?No.?1

64·论著·

作者单位： 复旦大学附属华山医院抗生素研究所，卫生计生委抗

生素临床药理重点实验室，上海 200040。

作者简介： 周迎（1993—），女，硕士研究生，主要从事细菌耐

药性及耐药机制研究。

通信作者：徐晓刚，E-mail ：xuxiaogang@https://www.360docs.net/doc/d318508444.html, 。临床分离万古霉素耐药肠球菌van 基因型及分子分型周?迎，?杨?洋，?郭?燕，?郑永贵，?吴?湜，?徐晓刚

摘要： 目的 明确临床分离万古霉素耐药肠球菌（VRE ）基因型及分子分型。方法 收集临床分离VRE 17株；采用16S rRNA 测序对临床株进行菌种确认，微量稀释法和琼脂稀释法测定常用抗菌药物最低抑菌浓度（MIC ）。多重PCR 进行van 基因分型；采用多位点序列分型（MLST ）进行分子分型。结果 17株VRE 经16S rRNA 测序确认均为屎肠球菌；其中12株对替考拉宁耐药。13株检出vanA 基因，9株检出vanM 基因，5株同时检出vanA 及vanM 基因；17株VRE 分属6个MLST 型，其中ST78型8株、ST80型4株、ST555型2株，其余3株分别为ST117型、ST262型和ST341型。结论 VRE 的临床分离株中van 基因型主要为vanA （76.5%），其次为vanM （52.9%），首次发现同时携带vanA 及vanM 的VRE 菌株。

关键词： 万古霉素； 耐药； 基因簇； 屎肠球菌

中图分类号：R978.1 文献标识码：A 文章编号：1009-7708 ( 2018 ) 01-0064-04

DOI: 10.16718/j.1009-7708.2018.01.013

van genotype and molecular typing of clinical isolates of vancomycin-resistant Enterococcus

ZHOU Ying, YANG Yang, GUO Yan, ZHENG Yonggui, WU Shi, XU Xiaogang. （Institute of Antibiotics, Huashan Hospital, Fudan University; Key Laboratory of Clinical Pharmacology of Antibiotics, National Population and Family Planning Commission, Shanghai 200040, China ）

Abstract: Objective To determine the genotype and molecular typing of vancomycin-resistant Enterococcus (VRE). Methods Seventeen clinical isolates of VRE were collected in 2016. The strains were identified to species and confirmed by 16S rRNA sequencing. The minimum inhibitory concentration of antimicrobial agents was determined by microdilution method and agar dilution method. Multilocus sequence typing (MLST) was used for molecular typing. Results The VRE strains were confirmed as Enterococcus faecium by 16S rRNA sequencing. All strains were resistant to vancomycin, but only 12 strains were resistant to teicoplanin. The vanA gene was identified in 13 of the 17 strains. The vanM gene was detected in 9 strains. Both vanA and vanM genes were identified in five of the 17 strains. Six MLST types were identified in the 17 strains, including ST78 (n =8), ST80 (n =4), ST555 (n =2), and one each for ST117, ST262 and ST341. Conclusions The van genotype was primarily vanA (76.5%) and vanM (52.9%) in clinical isolates of VRE. The VRE strains carrying both vanA and vanM were found for the first time.

Key words: vancomycin; resistance; gene cluster; Enterococcus faecium

万古霉素耐药肠球菌（vancomycin-resistant

Enterococcus ，VRE ）已经逐渐成为医院感染的重

要病原菌，可引起尿路、腹腔、伤口感染，以及

血流感染、心内膜炎等严重感染性疾病。中国大陆地区VRE 主要携带vanA 和vanM 两型耐药基因簇 [1-2]，vanA 与vanM 耐药基因簇均属于D-丙氨酸：D-乳酸连接酶（D-Ala ：D-Lac ligase ）基因簇，常介导宿主菌对万古霉素等糖肽类抗菌药物耐药。本研究对2016年分离自上海地区6所医院的17株VRE 进行耐药基因及多位点序列分型（MLST ）分析，为此类耐药菌感染防治提供参考。