CAJ | 학술논문

目的：探讨铜绿假单胞菌对喹诺酮类药物的耐药机制。方法30株对环丙沙星耐药的铜绿假单胞菌，采用 PCR方法扩增DNA解旋酶和拓扑异构酶Ⅳ的基因 gyrA，gyrB，parC和 parE，再测序查找是否存在位点突变，同时用脉冲场电泳（pulsed-field gel electrophoresis，PFGE）对菌株进行同源性分析。结果30株菌中，28株菌出现 gyrA基因扩增片段的137位点均有C→T突变，导致T83I改变；17株菌gyrB基因扩增片段的351位出现G→C突变，导致G466A改变；21株菌的 parC基因扩增片段的277位点有 C→U 突变导致 S87L 改变；2株菌 parE 基因在不同位点出现 C→U 突变，导致A425V和 A473V改变。30株菌可分为6个克隆，其中 A克隆4株，仅有 gyrA突变；B克隆7株，有 gyrA和 parC两种基因发生突变；C克隆3株，有gyrA和gyrB两种基因发生突变；D克隆14株，同时有 gyrA，gyrB和 parC三种基因发生突变；其他克隆2株，仅在 parE位点发生突变。结论该组菌株的靶位突变型与流行克隆型密切相关，同一流行型的菌株药物作用靶位的改变相同，并与环丙沙星的 MICs值的高低呈正比，突变基因数越多，MICs 值越高。4种基因中 gyrA基因突变频率最高，且该突变比其他靶位的突变对药物与靶位结合的影响更大，是需要关注的重点。
목적：탐토동록가단포균대규낙동류약물적내약궤제。방법30주대배병사성내약적동록가단포균，채용 PCR방법확증DNA해선매화탁복이구매Ⅳ적기인 gyrA，gyrB，parC화 parE，재측서사조시부존재위점돌변，동시용맥충장전영（pulsed-field gel electrophoresis，PFGE）대균주진행동원성분석。결과30주균중，28주균출현 gyrA기인확증편단적137위점균유C→T돌변，도치T83I개변；17주균gyrB기인확증편단적351위출현G→C돌변，도치G466A개변；21주균적 parC기인확증편단적277위점유 C→U 돌변도치 S87L 개변；2주균 parE 기인재불동위점출현 C→U 돌변，도치A425V화 A473V개변。30주균가분위6개극륭，기중 A극륭4주，부유 gyrA돌변；B극륭7주，유 gyrA화 parC량충기인발생돌변；C극륭3주，유gyrA화gyrB량충기인발생돌변；D극륭14주，동시유 gyrA，gyrB화 parC삼충기인발생돌변；기타극륭2주，부재 parE위점발생돌변。결론해조균주적파위돌변형여류행극륭형밀절상관，동일류행형적균주약물작용파위적개변상동，병여배병사성적 MICs치적고저정정비，돌변기인수월다，MICs 치월고。4충기인중 gyrA기인돌변빈솔최고，차해돌변비기타파위적돌변대약물여파위결합적영향경대，시수요관주적중점。
Objective 30 Pseudomonasaeruginosa mechanism of resistance to quinolones.Methods For the determination of ciprofloxacin MIC by agar dilution method.Used PCR on DNA gyrase and topoisomerase Ⅳ,resistance genes gyrA,gyrB, parC and parE were amplified,and BLAST,to determine whether there was resistance to bits mutation point;using pulsed-field gel electrophoresis (PFGE)of these 30 strains homology analysis.Results The 28 bacterial strains gyrA gene ampli-fied fragment of 137 points were C→T mutation causes T83I;17 strains gyrB gene amplified fragment of 351 G→C lead to G466A;parC gene amplification 21 bacteria fragment 277 point increase with C→U mutation causes S87L change two differ-ent strains parE gene locus C→U mutation A425V and A473V cause change.PFGE results:30 Pseudomonas aeruginosa could be divided into six clones,Aclone 4,B clone 7,C clone 3,D clone 14,and two other single clones.Conclusion The tar-get mutant strains closely related to the epidemic clone type,the same changes in the same pop-type strains of drug targets, and proportional to the level of ciprofloxacin MICs value,the more the number of mutated genes,MICs value higher.GyrA gene most prone to mutation,the mutation was also the first to be discovered,more than any other target of the mutation mutations on binding of drugs and targets that would be the focus of concern.