铜绿假单胞菌氨基糖苷类抗生素耐药基因检测

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:rmtB和ar-mA,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rmtB, armA.

Detection of aminoglycoside antibiotic resistance genes of Pseudomonas aeruginosa

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 16S rRNA methylase genes were detected by PCR,the positive gene amplification products were further verified by gene se-quencing. 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 detec-ted, 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 16S rRNA methylase gene were detec-ted, including rmtB and armA, with the positive rate of 29.6% and 29.6% , respectively, no other gene was de-tected. 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 (6ˊ)-Ⅰb, ant (2″) -Ⅰa, rmtB and armA, respectively.