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抗菌药联合外排泵抑制剂对铜绿假单胞菌耐药突变选择窗的影响

Effects of antibiotics plus efflux pump inhibitors on mutant selection window of Pseudomonas aeruginosa in vitro

摘要:

目的:探讨抗菌药联合外排泵抑制剂( EPI)对铜绿假单胞菌耐药突变选择窗的影响。方法选择铜绿假单胞菌标准株,测定美罗培南、头孢他啶、阿米卡星、环丙沙星4种药物单药最低抑菌浓度( MIC);棋盘格法测定联合利血平、奥美拉唑、阿奇霉素、羰基氰化物间氯苯腙( CCCP )、苯丙氨酸-精氨酸-β-萘胺(PAβN)5种EPI抗菌药物的MIC。琼脂平板二倍稀释法测定抗菌药物单药以及联合EPI的防耐药突变浓度(MPC),计算相应的选择指数(MPC/MIC);分别测定美罗培南、头孢他啶、环丙沙星联合阿米卡星,阿米卡星联合环丙沙星后的选择指数。在临床株中检测美罗培南、头孢他啶、阿米卡星、环丙沙星单药及与CCCP联合后的MIC、MPC及选择指数。结果在标准株,美罗培南、头孢他啶、阿米卡星、环丙沙星单药的选择指数分别为>32,>32,16,16;与利血平联合后选择指数分别为16、16、16、8;与奥美拉唑联合后分别为16、32、16、8;与阿奇霉素联合后分别为8、16、16、8;与CCCP联合后分别为8、8、16、8;与PAβN联合后分别为8、16、16、8。美罗培南、头孢他啶、环丙沙星分别与阿米卡星联合后的选择指数为4、4、8。阿米卡星与环丙沙星联合后阿米卡星的选择指数为4。在临床株,环丙沙星联合CCCP后选择指数较单药显著降低。结论除阿米卡星外,美罗培南、头孢他啶、环丙沙星联合EPI可明显降低MIC及MPC,环丙沙星联合CCCP可缩小突变选择窗,有利于减少耐药突变体的产生。

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abstracts:

Objective To explore the effects of antibiotics plus efflux pump ( EPI ) inhibitors on mutant selection window of Pseudomonas aeruginosa in vitro.Methods In standard strains , the minimum inhibitory concentrations ( MICs ) of meropenem , ceftazidime , amikacin and ciprofloxacin alone were measured.Then the MICs of four antibiotics respectively plus EPI , including reserpine , omeprazole , azithromycin, carbonyl cyanide m-chlorophenyl-hydrazone (CCCP) and Phe-Arg-β-naphtylamide (PAβN), were measured by checkerboard test.And the mutant prevention concentrations ( MPC) of antibiotics alone and antibiotics plus different EPIs were also measured by agar dilution method.Selection indices ( SI) were acquired through MPC divided by MIC.Similarly, the SI of meropenem , ceftazidime and ciprofloxacin which respectively plus amikacin and amikacin plus ciprofloxacin were measured.In clinical strains, MIC, MPC and SI of antibiotics alone and four antibiotics plus CCCP were measured.Results In standard strains , the SI of meropenem, ceftazidime, amikacin and ciprofloxacin alone were >32, >32, 16 and 16.After adding EPI, the SI of four antibiotics were 16, 16, 16, 8 (reserpine), 16, 32, 16, 8 (omeprazole), 8, 16, 16, 8 (azithromycin), 8, 8, 16, 8 (CCCP) and 8, 16, 16, 8 (PAβN).The SI of meropenem, ceftazidime and ciprofloxacin plus amikacin were 4, 4, 8.And the SI of amikacin plus ciprofloxacin was 4.In clinical strains, the SI of ciprofloxacin significantly decreased after adding CCCP.Conclusions Both MIC and MPC of meropenem , ceftazidime and ciprofloxacin decreased significantly after adding EPI.The mutant selection window decreased when ciprofloxacin was combined with CCCP.

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