耐阿米卡星铜绿假单胞菌氨基糖苷类耐药基因及其基因型研究  

The study on aminoglycoside resistance gene and its genotyping of amikacin-resistant mechanism in Pseudomonas aeruginosa

作　　者：由然[1] 孙立颖[1] 

机构地区：[1]北京大学第一医院检验科,北京100034

出　　处：《检验医学》2010年第5期341-344,共4页Laboratory Medicine

摘　　要：目的了解耐阿米卡星(AMK)铜绿假单胞菌(Pa)的氨基糖苷类修饰酶基因和氨基糖苷耐药基因的存在情况及其基因型。方法对临床分离的72株多重耐药Pa(其中对AMK耐药Pa组和对AMK敏感Pa组各36株)分别用聚合酶链反应(PCR)测定9种主要的氨基糖苷类修饰酶基因和2种氨基糖苷耐药基因,用基因外重复回文序列(REP)-PCR进行基因型研究。结果在分组研究中AMK耐药Pa组表现为aac(6′)-Ⅰ和aac(3)-Ⅱ基因同时阳性的占42.4%,aac(6′)-Ⅰ和ant(2″)-Ⅰ基因同时阳性的占36.4%,是耐药组的主要修饰酶基因;AMK敏感Pa组表现为aac(6′)-Ⅰ基因阳性的占50.0%,aac(3)-Ⅱ基因阳性的占33.2%,是敏感组的主要修饰酶基因。2种氨基糖苷耐药基因(RmtA和RmtD)均为阴性。REP-PCR显示耐药菌株可分为A～G 7型,其中AMK耐药Pa组的主要分型为F型(其中尤以F1型为多);AMK敏感Pa组以E型为多见。结论通过2种或多种氨基糖苷类修饰酶同时作用于AMK是Pa对AMK耐药的重要途径之一。Objective To study the existing situation and genotyping of aminoglycoside modifying enzyme gene and aminoglycoside resistance gene of amikacin（AMK）-resistant mechanism in Pseudomonas aeruginosa（Pa）.Methods 72 strains of mutli-resistant Pa,including 36 AMK-resistant Pa and 36 AMK-sensitive Pa,were detected for 9 main kinds of aminoglycoside modifying enzyme gene and 2 aminoglycoside resistance genes by polymerase chain reaction（PCR）.The genotyping was analyzed and investigated by repetitive extragenic palindromic（REP）-PCR.Results In AMK-resistant Pa group,there was 42.4% for aac（6′）-Ⅰand aac（3）-Ⅱ genes both positive,and there was 36.4% for aac（6′）-Ⅰand ant（2″）-Ⅰgenes both positive which were the main modifying enzyme genes in resistant group.In AMK-sensitive Pa group,aac（6′）-Ⅰgene positive was 50.0%,and aac（3）-Ⅱgene positive was 33.2%.They were the main modifying enzyme genes in sensitive group.2 kinds of aminoglycoside resistance genes（RmtA gene and RmtD gene） were not found.The results of REP-PCR showed that there were 7 types from A to G.In AMK-resistant Pa group,the main genotyping was F（especially F1）.In AMK-sensitive Pa group,the main genotyping was E.Conclusions It′s one of significant ways of AMK-resistant mechanism in Pa that two or more than two aminoglycoside modifying enzymes act on AMK simultaneously.

关 键 词：铜绿假单胞菌 阿米卡星 氨基糖苷类修饰酶基因 RmtA基因 RmtD基因 基因外重复回文序列-聚合酶链反应 

分 类 号：R378.99[医药卫生—病原生物学]