临床铜绿假单胞菌对头孢他啶异质性耐药研究  

Investigation of Ceftazidime Heteroresistance in Pseudomonas aeruginosa Clinical Isolates

作　　者：李金玲 李文茹 张建设 黄旭斌[3] 廖康[4] 谢小保 LI Jin-ling;LI Wen-ru;ZHANG Jian-she(National Engineering Research Center for Marine Aquaculture of Zhejiang Ocean University,Zhoushan 316022;Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application,State Key Laboratory of Applied Microbiology Southern China,Institute of Microbiology,Guangdong Academy of Sciences,Guangzhou 510070,China)

机构地区：[1]浙江海洋大学国家海洋设施养殖工程技术研究中心,浙江舟山316022 [2]广东省科学院微生物研究所,华南应用微生物国家重点实验室,广东省菌种保藏与应用重点实验室,广东广州510070 [3]中山大学附属第一医院呼吸与危重症医学科,广东广州510080 [4]中山大学附属第一医院医学检验科,广东广州510080

出　　处：《浙江海洋大学学报（自然科学版）》2022年第4期357-365,共9页Journal of Zhejiang Ocean University：Natural Science

基　　金：广东省基础与应用基础研究基金(2021A1515011080);国家自然科学基金(31500113)。

摘　　要：初步探究了铜绿假单胞菌对头孢他啶的异质性耐药情况及其特征,为临床感染治疗提供参考。本研究收集了临床分离的50株铜绿假单胞菌,通过药敏纸片扩散法(Kirby-Bauer,K-B)、菌落谱型分析法(population analysis profiling,PAP)、适应成本测定以及耐药稳定性实验探究铜绿假单胞菌的异质性耐药特征。K-B结果显示铜绿假单胞菌对头孢他啶(CAZ)异质性耐药率为40%(20/50),经过PAP实验确认后有4株是异质性耐药菌株,占总菌株的8%(4/50),其耐药亚群的发生频率为1×10^(-7)~1.2×10^(-5)。适应性成本测定结果表明,异质性耐药菌株PAS56的3株耐药亚群的生长速率与其亲本菌株没有统计学差异(P>0.05),而PAS95的3株耐药亚群却比亲本菌株生长速率更快,有极显著差异(P<0.01)。从PAS56和PAS95的PAP实验32μg·mL^(-1)平板上各挑取了3株耐药亚群分离株,并进行了无抗生素压力下的传代培养,耐药亚群的耐药性能稳定遗传,且均高于原始菌株水平。铜绿假单胞菌对头孢他啶的异质性耐药率较低,但其耐药亚群发生频率相对较高,并且耐药性稳定,极易转化为高水平耐药菌株。因此在使用头孢他啶进行临床治疗时应更加注意异质性耐药的发生,需采用有效的检测方法,为临床防控和合理使用抗生素提供指导。The heteroresistance and characteristics of Pseudomonas aeruginosa were investigated in this study to provide reference for clinical infection treatment.Fifty strains of P.aeruginosa were collected from clinical isolates,and their heteroresistance characteristics were studied by Kirby-Bauer(K-B),population analysis profiling(PAP),fitness cost and resistance stability assay.The heteroresistance rate of P.aeruginosa to ceftazidime(CAZ)was 40%(20/50)by K-B primary screening.Four strains were confirmed to be heteroresistance by PAP,accounting for 8%(4/50)of the total strains,and the frequency of their resistant subgroups was 1×10^(-7)~1.2×10^(-5).Fitness cost experimental results showed that there was no statistical difference in the growth rate between three resistant subpopulations and parental strain of the PAS56 heteroresistance strain(P>0.05),while there was a very significant difference in the growth rate between three resistant subpopulations and parental strains of the PAS95 heteresistance strain(P<0.01).The high-level resistant subpopulations picked from 32μg·mL^(-1) plate in the PAP experiments of PAS56 and PAS95 were cultured under no antibiotic pressure for 50 generations.The experimental results showed that resistance activities of all the heteresistance subpopulations were stable and their resistant levels were higher than that of the parental strains.The resistance subpopulation of P.aeruginosa to ceftazidime occurred at a relatively high frequency,though the heteroresistance rate was low.Moreover,the resistance levels of the heteroresistance subpopulation were high and stable,and then these heteroresistance subpopulations were easily transformed into a high level of resistant strains.Therefore,more attention should be paid to the occurrence of heteroresistance when using ceftazidime for P.aeruginosa clinical treatment.

关 键 词：铜绿假单胞菌 异质性耐药 头孢他啶 

分 类 号：Q599[生物学—生物化学] R978.11[医药卫生—药品]