检验医学
檢驗醫學
검험의학
LABORATORY MEDICINE
2014年
6期
646-650
,共5页
张国栋%王莹%朱红胜
張國棟%王瑩%硃紅勝
장국동%왕형%주홍성
铜绿假单胞菌%耐药机制%美罗培南%环丙沙星%外排系统
銅綠假單胞菌%耐藥機製%美囉培南%環丙沙星%外排繫統
동록가단포균%내약궤제%미라배남%배병사성%외배계통
Pseudomonas aeruginosa%Drug resistance mechanism%Meropenem%Ciprofloxacin%Efflux system
目的：探讨临床分离的对美罗培南和环丙沙星均耐药的铜绿假单胞菌的耐药机制。方法收集经VITEK-2 Compact微生物分析系统检测为美罗培南和环丙沙星耐药的铜绿假单胞菌30株，琼脂稀释法复查最小抑菌浓度（MIC）值。改良三维试验检测碳青霉烯酶，聚合酶链反应（PCR）扩增耐药基因，逆转录PCR 分析细菌外排系统表达情况。结果琼脂稀释法与VITEK-2 Compact微生物分析系统检测结果相同。所有菌株均未产碳青霉烯酶，PCR扩增测序未发现基因改变，逆转录PCR检测外排系统发现以mexX和mexC基因表达增加为主，其调控基因扩增测序发现4株mexC过度表达的调控基因nfxB Gly→Val（GGC→GTC），6株mexX过度表达的调控基因mexZ Val→Gly（CTG→CAG）。结论外排系统调控基因突变而引起的外排系统MexXY-OprM和MexCD-OprJ过度表达是引起美罗培南和环丙沙星耐药的主要原因。
目的：探討臨床分離的對美囉培南和環丙沙星均耐藥的銅綠假單胞菌的耐藥機製。方法收集經VITEK-2 Compact微生物分析繫統檢測為美囉培南和環丙沙星耐藥的銅綠假單胞菌30株，瓊脂稀釋法複查最小抑菌濃度（MIC）值。改良三維試驗檢測碳青黴烯酶，聚閤酶鏈反應（PCR）擴增耐藥基因，逆轉錄PCR 分析細菌外排繫統錶達情況。結果瓊脂稀釋法與VITEK-2 Compact微生物分析繫統檢測結果相同。所有菌株均未產碳青黴烯酶，PCR擴增測序未髮現基因改變，逆轉錄PCR檢測外排繫統髮現以mexX和mexC基因錶達增加為主，其調控基因擴增測序髮現4株mexC過度錶達的調控基因nfxB Gly→Val（GGC→GTC），6株mexX過度錶達的調控基因mexZ Val→Gly（CTG→CAG）。結論外排繫統調控基因突變而引起的外排繫統MexXY-OprM和MexCD-OprJ過度錶達是引起美囉培南和環丙沙星耐藥的主要原因。
목적：탐토림상분리적대미라배남화배병사성균내약적동록가단포균적내약궤제。방법수집경VITEK-2 Compact미생물분석계통검측위미라배남화배병사성내약적동록가단포균30주，경지희석법복사최소억균농도（MIC）치。개량삼유시험검측탄청매희매，취합매련반응（PCR）확증내약기인，역전록PCR 분석세균외배계통표체정황。결과경지희석법여VITEK-2 Compact미생물분석계통검측결과상동。소유균주균미산탄청매희매，PCR확증측서미발현기인개변，역전록PCR검측외배계통발현이mexX화mexC기인표체증가위주，기조공기인확증측서발현4주mexC과도표체적조공기인nfxB Gly→Val（GGC→GTC），6주mexX과도표체적조공기인mexZ Val→Gly（CTG→CAG）。결론외배계통조공기인돌변이인기적외배계통MexXY-OprM화MexCD-OprJ과도표체시인기미라배남화배병사성내약적주요원인。
Objective To investigate the drug resistance mechanisms of both meropenem and ciprofloxacin in clinical isolates of Pseudomonas aeruginosa.Methods A total of 30 isolates of Pseudomonas aeruginosa which had been tested resistance to both meropenem and ciprofloxacin by VITEK-2 Compact were collected.Agar dilution method was used to reexamine the minimal inhibitory concentrations (MIC)of meropenem and ciprofloxacin.Carbapenemases were detected by modified three-dimensional test.The resistance genes were analyzed by polymerase chain reaction (PCR) amplification.The expression of efflux systems was analyzed by reverse transcription PCR.Results The same results were showed between agar dilution method and VITEK-2 Compact.Carbapenemases were not detected by modified three-dimensional test.The overexpression of mexX and mexC genes were detected by reverse transcription PCR.The sequencing of regulatory genes showed that there were 4 isolates which the Gly(GGC)of nfxB was substitute for Val (GTC)and 6 isolates which the Val(CTG)of mexZ was substitute for Gly (CAG).Conclusions The overexpression of the MexXY-OprM and MexCD-OprJ efflux system,due to the mutation of regulatory genes of efflux system,is contributed to the drug resistance to both meropenem and ciprofloxacin of Pseudomonas aeruginosa.
