耐亚胺培南鲍曼不动杆菌主动外排系统AdeIJK的表达研究  

Study on expression of active efflux system AdeIJK involved in imipenem-resistant Acinetobacter baumannii

作　　者：潘双[1] 刘原[1] 柯蕊[1] 和平[1] 李满祥[1] 杨芬[1] 耿燕[2] 张毅[2] 

机构地区：[1]西安交通大学医学院第二附属医院呼吸内科,710004 [2]西安交通大学医学院第二附属医院检验科,710004

出　　处：《国际呼吸杂志》2014年第22期1681-1685,共5页International Journal of Respiration

基　　金：国家自然科学基金（81270061）;陕西省科技攻关项目（2010K1601（2））

摘　　要：目的调查鲍曼不动杆菌耐药性，研究亚胺培南耐药性与主动外排系统AdeIJK的相关性。方法收集非重复性鲍曼不动杆菌共362株，K—B法和微量肉汤稀释法分别进行体外药敏试验；采用PCR法扩增主动外排系统基因adeI、adeJ、adeK；采用RT—PCR法对亚胺培南敏感菌株和耐药菌株中的adeJ基因进行扩增，凝胶扫描半定量分析adeJ基因在mRNA水平的表达情况。结果73株鲍曼不动杆菌对亚胺培南耐药，耐药率为20．17％，亚胺培南耐药菌株对头孢哌酮／舒巴坦耐药性最低，为45．21％；PCR法检测到鲍曼不动杆菌adeJ基因的检出率为93．89％，在检出adeJ基因的菌株中有3株未检出adeI基因，均检出adeK基因；RT—PCR法检测到12株亚胺培南敏感菌株和5株耐药菌株的adeJ基因相对表达水平（灰度）分别为0．3485±4-0．2671、0．8999±4-0．2773，统计学分析显示：adeJ基因在敏感组和耐药组之间的表达存在差异性。结论耐亚胺培南鲍曼不动杆菌呈现多重耐药和泛耐药趋势；耐亚胺培南鲍曼不动杆菌的耐药性可能与AdeIJK的高表达有关。Objective To investigate the antimicrobial resisitance of Acinetobacter baumannii in Xi＇an area,and to study the effection of the efflux pump system AdeIJK involved in irnipenern-resistant Acinetobacter baurnannii. Methods 362 non-repeatability Acinetobacter baumannii strains were collected. The K-B method and broth mierodilution method were applied to detect drug sensitivity of all strains. Active efflux system genes adeI,adeJ,adeK were amplified by polyrnerase chain reaction （PCR）. The adeJ gene in imipenem-resistant strains and irnipenern-sensitive strains were amplified by reverse transcription polyrnerase chain reaction （RT-PCR）, the expression of adeJ mRNA was analyzed by semiquantitative analysis of gel electrophoresis. Results 73 non-repetitive imipenem-resistant Acinetobacter baurnannii strains were selected,the resistance rate to imipenern was 20.17%, the resistance rate of all imipenem- resistant Aeinetobacter baurnannii strains to cefoperazone/sulbactarn was 45.21% （the lowest resistance rate）. By PCR technology, the positive rate of adeJ gene in Acinetobaeter baurnannii was 93.89%, adeI gene was not detected in three adeJ-positive strains,adeK gene was detected in all adeJ-positive strains. By RT-PCR technology, the relative intensity of rnRNA expression of adeJ was 0. 348 5 ±0. 267 1 and 0. 899 94±0. 277 3 in 12 imipenem-sensitive strains and five imipenern-resistant strains. Statistical analysis showed the expression of adeJ was difference between the sensitive group and the resistant group.Conclusions The imipenem-resistant Acinetobacter baumannii strains in our region shows the trend ofmultidrug-resistance and pan-drug resistance. The resistance o{ the imipenem-resistant Acinetobacter baumannii may be related to the overexpression of active ef{lux system AdeIJK.

关 键 词：鲍曼不动杆菌 亚胺培南 多重耐药 主动外排系统AdeIJK RT PCR 

分 类 号：R446.5[医药卫生—诊断学]