铜绿假单胞菌氨基糖苷类抗生素耐药基因检测  

Detection of aminoglycoside antibiotic resistance genes of Pseudomonas aeruginosa

作　　者：张凡[1,2] 郭普[2] 张伦军[2] 郑晶[2] 郑照军[2] 徐元宏[1] Zhang Fan;Guo Pu;Zhang Lunjun(Dept of Clinical Laboratory,The First Affiliated Hospital of Anhui Medical University,Hefei 230022;Dept of Clinical Laboratory,The First Affiliated Hospital of Bengbu Medical College,Bengbu 233004)

机构地区：[1]安徽医科大学第一附属医院检验科,合肥230022 [2]蚌埠医学院第一附属医院检验科,蚌埠233004

出　　处：《安徽医科大学学报》2018年第6期928-932,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省高校自然科学研究项目(编号:KJ2015B044by)

摘　　要：目的了解铜绿假单胞菌对氨基糖苷类抗生素耐药基因的分布情况。方法收集临床分离的铜绿假单胞菌,采用仪器法和纸片扩散法(K-B法)进行药敏实验。筛选出对氨基糖苷类抗生素耐药的菌株,用PCR法检测16个氨基糖苷类修饰酶(AMEs)基因和5个16S rRNA甲基化酶基因的表达,进一步对阳性基因扩增产物进行测序验证。结果 54株菌中51株AMEs基因阳性,检出率为94.4%,28株甲基化酶基因阳性,检出率为51.9%。共检出5种AMEs基因:ant(3″)-Ⅰ、aac(3)-Ⅱc、ant(4')-Ⅰa、aac(6')-Ⅰb和ant(2″)-Ⅰa,阳性率分别为66.7%、38.9%、31.5%、31.5%和16.6%;2种16S rRNA甲基化酶基因:rmt B和arm A,阳性率分别为29.6%和29.6%,其余基因均未检出。测序结果与目的基因一致。结论铜绿假单胞菌氨基糖苷类抗生素耐药的主要基因为ant(3″)-Ⅰ、aac(3)-Ⅱc、ant(4')-Ⅰa、aac(6')-Ⅰb和ant(2″)-Ⅰa以及rmt B,arm A。Objective To investigate the aminoglycoside antibiotic resistance gene distribution of Pseudomonas aeruginosa. Methods Pseudomonas aeruginosa was isolated and collected in clinic. Drug susceptibility test was performed by instrumental method and disk diffusion method（ K-B method）. Aminoglycoside antibiotic-resistant strains were screened,and the expression of 16 aminoglycoside modified enzyme（ AMEs） genes and 5 16 S rRNA methylase genes were detected by PCR,the positive gene amplification products were further verified by gene sequencing. Results Among the 54 strains,51 were positive for AMEs gene,with the detection rate of 94. 4%; 28 strains were positive for methylase gene,with the detection rate of 51. 9%. Five kinds of AMEs gene were detected,including ant（ 3″）-Ⅰ,aac（ 3）-Ⅱc,ant（ 4’）-Ⅰa,aac（ 6’）-Ⅰb and ant（ 2″）-Ⅰa,with the positive rate of 66. 7%,38. 9%,31. 5%,31. 5% and 16. 6% respectively; Two kinds of 16 S rRNA methylase gene were detected,including rmt B and arm A,with the positive rate of 29. 6% and 29. 6%,respectively,no other gene was detected. The gene sequencing result was consistent with the target gene sequence. Conclusion The aminoglycoside resistance gene of Pseudomonas aeruginosa was detected with the 5 types of ant（ 3″）-Ⅰ,aac（ 3）-Ⅱc,ant（ 4’）-Ⅰa,aac（ 6’）-Ⅰb,ant（ 2 ＂）-Ⅰa,rmt B and arm A,respectively.

关 键 词：铜绿假单胞菌 耐药基因 氨基糖苷类修饰酶 甲基化酶 

分 类 号：R378.991[医药卫生—病原生物学]