中国临床药理学与治疗学2023，Vol.28Issue(12)：1347-1356,10.DOI:10.12092/j.issn.1009-2501.2023.12.004
阿奇霉素联合左氧氟沙星对不同ST型肺炎克雷伯菌生物膜的抑制和清除作用的研究
Inhibitory and clearance effect of azithromycin combined with levo-floxacin on biofilm of different ST types of Klebsiella pneumoniae
摘要
Abstract
AIM:Given the biofilm formation abili-ty of different ST-type Klebsiella pneumoniae,our study was aimed at exploring the inhibitory and clearance of azithromycin combined with levofloxa-cin on the biofilm of Klebsiella pneumoniae of dif-ferent ST-types and providing a new strategy for the prevention of biofilm formation in the treat-ment of post-infectious Klebsiella pneumoniae.METHODS:9 strains of Klebsiella pneumoniae from all susceptibility groups,19 strains of Klebsiella pneumoniae producing extended-spectrum β-lac-tamases(ESBLs),and 37 strains of Carbapenem-re-sistant Klebsiella pneumoniae(CRKP)were random-ly collected from the samples of patients hospital-ized in the First Affiliated Hospital of Wannan Medi-cal College from August 2019 to November 2021.The isolates were identified using VITEK MS IVD KB V3.2 and VITEK 2-Compact 60.Multilocus sequence typing(MLST)was performed to analyze the homol-ogy of each strain;crystal violet staining was used for semi-quantitative detection of biofilm to com-pare the differences in biofilm formation ability be-tween different ST-type Klebsiella pneumoniae.Dif-ferent ST-type strains were selected,and the par-tial inhibitory concentration index(FICI)was calcu-lated by micro broth dilution method to judge the combination effect and select the optimal combina-tion concentration;crystalline violet staining meth-od was used to investigate the inhibition and clear-ance effect of azithromycin combined with levoflox-acin on the biofilm of different ST-type Klebsiella pneumoniae;laser scanning confocal fluorescence microscopy was used to observe the structural changes of the biofilm of Klebsiella pneumoniae be-fore and after the effect of the antibacterial drugs.RESULTS:MLST typing results showed that the sen-sitive group of Klebsiella pneumoniae strains had 8 sequences such as ST86,ST727,etc.,the ESBLs group strains belonged to 14 sequence types such as ST15,ST37,ST11,etc.,of which ST15 accounted for 26.32％(5/19).The CRKP group strains be-longed to 9 sequence types such as ST11,ST15,ST656,etc.,of which ST11 accounted for 48.65％(18/37),ST15 accounted for 27.03％(10/37);ST15(ESBLs),ST11(CRKP),ST15(CRKP)type Klebsiella pneumoniae biofilms all reached maturity on the 5th day,the ST15(ESBLs)group had a stronger abil-ity to produce material to be membranous than the ST15(CRKP)group.The ST11(CRKP)group had a stronger ability to produce material to be mem-branous than the ST15(CRKP)group(P＜0.01);the results of azithromycin combined with levofloxacin drug sensitivity showed that it had an additive ef-fect on different ST-type Klebsiella pneumoniae bacteria;in the inhibition of biofilm formation and clearance test,the 2×MIC azithromycin group and the combined concentration group had a stronger inhibition of biofilm formation of different ST-type Klebsiella pneumoniae bacteria,and the inhibitory ability of the combined group was better than that of the single-drug group,and the highest inhibition rate could reach 89.93％;the clearance effects were all combined drug group＞azithromycin＞levo-floxacin,and the highest clearance rate was 44.79％.CONCLUSION:There are differences in bio-film formation ability between different ST-type Klebsiella pneumoniae,and azithromycin combined with levofloxacin has a better inhibitory effect on different ST-type Klebsiella pneumoniae biofilm,conbined application can be used in the treatment of biofilm infections early stage.关键词
肺炎克雷伯菌/阿奇霉素/左氧氟沙星/多位点序列分型/生物膜Key words
Klebsiella pneumoniae/azithromycin/levofloxacin/multilocus sequence typing/biofilm分类
医药卫生引用本文复制引用
曹若楠,李小宁,阮欣然,刘诗惠,陈婧璇,徐浩,沈继录,朱国萍..阿奇霉素联合左氧氟沙星对不同ST型肺炎克雷伯菌生物膜的抑制和清除作用的研究[J].中国临床药理学与治疗学,2023,28(12):1347-1356,10.基金项目
2020年安徽省属本科医学院校公共卫生协同创新项目(GXXT-2020-016) （GXXT-2020-016）
分子酶学与重大疾病机理研究安徽省重点实验室开放课题(fzmx202015) （fzmx202015）
2020年度安徽高校自然研究项目(KJ2020A0610) （KJ2020A0610）
