铜绿假单胞菌临床分离株新型氨基糖苷类修饰酶基因分析  

New aminoglycoside modifying enzyme gene in clinical isolates of Pseudomonas aeruginosa

作　　者：朱健铭 翁幸鐾[2] 姜如金 吴晋兰[4] 凌莉萍[1] ZHU Jian-ming;WENG Xing-bei;JIANG Ru-jin;WU Jin-lan;LING Li-ping(Hangzhou Yuhang Hospital of Traditional Chinese Medicine,Hangzhou,Zhejiang 311106,China;不详)

机构地区：[1]杭州市余杭区中医院检验科,浙江杭州311106 [2]宁波市第一医院检验科,浙江宁波315010 [3]杭州市余杭区第一人民医院检验科,浙江杭州311100 [4]杭州市余杭区第五人民医院中医内科,浙江杭州311100

出　　处：《中华医院感染学杂志》2021年第15期2251-2255,共5页Chinese Journal of Nosocomiology

基　　金：浙江省医药卫生科技计划基金资助项目(2019KY552);杭州市科委医疗卫生科研基金资助项目(2018HZ001);杭州市余杭区医疗卫生重大科研基金资助项目(2018YH001)。

摘　　要：目的了解铜绿假单胞菌临床分离株对氨基糖苷类药物耐药的遗传学背景。方法收集2018年3月-2019年5月浙江省宁波与杭州两个地区6所医院的耐药铜绿假单胞菌98株(非重复株),采用琼脂稀释法测定其对13种抗菌药物的最低抑菌浓度(MIC);聚合酶链反应检测14种氨基糖苷类修饰酶基因和10种16S rRNA甲基化酶基因;比较宁波与杭州地区分离株耐药率和耐药基因携带率。结果98株铜绿假单胞菌对阿米卡星、妥布霉素和庆大霉素3种氨基糖苷类药物耐药率均<30%,对其余10种抗菌药物的耐药率高达60.20%~94.90%。宁波分离株对3种氨基糖苷类药物的耐药率均高于杭州分离株。98株菌共检出5种氨基糖苷类修饰酶基因:ant(3'')-Ⅰ(13/98,13.27%)、ant(2'')-Ⅰ(11/98,11.22%)、aac(6')-Ⅰb(7/98,7.14%)、aph(3')-ⅩⅤ(4/98,4.08%)、aac(6')-Ⅱ(3/98,3.06%),和1种16S rRNA甲基化酶基因:rmtB(10/98,10.20%)。宁波分离株氨基糖苷类耐药基因ant(2'')-Ⅰ、ant(3'')-Ⅰ、aph(3')-ⅩⅤ和rmtB检出率高于杭州分离株。25株菌检出氨基糖苷类耐药基因(25/98,25.51%),其中15株携带2种及以上耐药基因,10株携带1种耐药基因。10株检出16S rRNA甲基化酶基因rmtB的菌株对氨基糖苷类药物的MIC均≥256μg/ml。耐药基因携带模式共有6种,与氨基糖苷类药物耐药表型相符。结论携带ant(3'')-Ⅰ、ant(2'')-Ⅰ、aac(6')-Ⅰb、aph(3')-ⅩⅤ、aac(6')-Ⅱ、rmtB基因是本组铜绿假单胞菌对氨基糖苷类药物耐药的主要原因。OBJECTIVE To investigate genetic background of clinical isolate strain of Pseudomonas aeruginosa(P.aeruginosa)’s drug resistance to aminoglycosides.METHODS From Mar 2018 to May 2019,98 non-repetitive strains of P.aeruginosa were collected from 6 hospitals in Ningbo and Hangzhou of Zhejiang Province in China.The minimum inhibitory concentrations(MICs)of P.aeruginosa to 13 antimicrobial agents were performed by agar dilution method;14 aminoglycoside modifying enzyme genes and 1016 S rRNA methylase genes were detected by PCR assay;the drug resistance rates and positive gene rates were compared between strains isolated from Ningbo and Hangzhou.RESULTS The drug resistance rates of 98 P.aeruginosa strains to 3 aminoglycosides,such as amikacin,tobramycin and gentamicin,were<30%,and the drug resistance rates to other 10 antimicrobial agents were 60.20%~94.90%.The drug resistance rates of P.aeruginosa strains to 3 aminoglycosides isolated from Ningbo to were significantly higher than strains isolated from Hangzhou.A total of 5 aminoglycoside modifying enzyme genes,including ant(3'')-Ⅰ(13/98,13.27%),ant(2'')-Ⅰ(11/98,11.22%),aac(6')-Ⅰb(7/98,7.14%),aph(3')-ⅩⅤ(4/98,4.08%),aac(6')-Ⅱ(3/98,3.06%),and 116 S rRNA methylase gene of rmtB(10/98,10.20%)were positive-detected.The positive-detection rate of ant(2'')-Ⅰ,ant(3'')-Ⅰ,aph(3')-ⅩⅤand rmtB genes in P.aeruginosa strains isolated in Ningbo were significantly higher than those in Hangzhou.Aminoglycoside resistant genes were positive in 25 strains(25/98,25.51%),of which 15 strains carring 2 or more drug resistant genes while 10 strains harboring only 1 drug resistant gene.The MICs of 10 strains harboring 16 S rRNA methylase gene rmtB to aminoglycosides were≥256μg/ml.There are 6 modes of drug resistant genes,which were consistent with aminoglycosides phenotypes.CONCLUSION P.aeruginosa strains carring ant(3'')-Ⅰ,ant(2'')-Ⅰ,aac(6')-Ⅰb,aph(3')-ⅩⅤ,aac(6')-Ⅱ,and rmtB genes played a key role in drug resistance to aminoglycosides.

关 键 词：铜绿假单胞菌 氨基糖苷类 耐药 氨基糖苷类修饰酶 16S rRNA甲基化酶 基因 

分 类 号：R378[医药卫生—病原生物学]