基于MALDI-TOF MS定量快速药敏试验在肺炎克雷伯菌对亚胺培南和替加环素药敏试验中的适用性分析  

Applicability analysis of rapid antimicrobial susceptible testing based on MALDI-TOF MS technique in imipenem and tigecycline susceptibility test of Klebsiella pneumoniae

作　　者：张龙桃 张安然 陈倩倩 孔海芳 胡志东[1] ZHANG Longtao;ZHANG Anran;CHEN Qianqian;KONG Haifang;HU Zhidong(Department of Medical Laboratory,General Hospital,Tianjin Medical University,Tianjin 300052,China;Department of Pathology,Women and Children′s Hospital of Qingdao,Qingdao266034,China)

机构地区：[1]天津医科大学总医院医学检验科,天津300052 [2]山东省青岛市妇女儿童医院病理科,青岛266034

出　　处：《天津医科大学学报》2024年第3期232-238,共7页Journal of Tianjin Medical University

摘　　要：目的:探讨基于基质辅助激光解吸电离飞行时间质谱法(MALDI-TOF MS)的定量快速药敏试验(rAST)在肺炎克雷伯菌(KP)对亚胺培南(IPM)和替加环素(TGC)药敏试验中的实用性分析。方法:选取天津医科大学总医院2018年1月至2021年6月120株临床非重复KP,应用微量肉汤稀释法(BMD)检测其对IPM和TGC的最低抑菌浓度(MIC)值。采用优化甲酸提取法提取KP菌体蛋白,进行基于基质辅助激光解吸电离飞行时间质谱法的快速药敏试验[(MALDI-TOF MS)-based rAST]检测IPM和TGC的MIC值,并评估最佳的菌液稀释度和孵育时间。采用Cohen′s kappa统计来评估(MALDI-TOF MS)-based rAST法和标准参考BMD的一致性。结果:在不同的孵育时间,上述两种方法处理后的质谱均能成功鉴定KP(P>0.05);但优化后的甲酸提取法简化了操作流程,并且在菌量较低情况下的成功鉴定率高于常规方案。稀释25倍即菌液含量为6×10^(6) CFU/mL、孵育时间分别为2 h和3 h条件下,(MALDI-TOF MS)-based rAST法测得IPM(P<0.01)和TGC(P<0.001)的MIC值效果最佳;此条件下,(MALDI-TOF MS)-based rAST与BMD测得的MIC值相当,分别为IPM 70.83%(85/120)、TGC 66.67%(40/60),基本一致性(EA)分别为98.33%(118/120)、98.33%(59/60)。结论:(MALDI-TOF MS)-based rAST缩短了报告时间,是一种可靠的快速定量药敏试验方法。Objective:To investigate the practicability of rapid antimicrobial susceptible testing(rAST)based on mass-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS)in the susceptibility test of Klebsiella pneumoniae(KP)to imipenem(IPM)and tigecycline(TGC).Methods:A total of 120 clinical non-repetitive KP strains from Tianjin Medical University General Hospital from January 2018 to June 2021 were selected,and the minimum inhibitory concentrations(MIC)of IPM and TGC were detected by broth microdilution(MIC).The optimal formic acid extraction method was used to extract the protein of KP.The MIC values of IPM and TGC were detected by rapid antimicrobial susceptible testing based on mass-assisted laser desorption ionization time-of-flight mass spectrometry[(MALDI-TOF MS)-based rAST],and the optimal dilution and incubation time of the bacterial solution were evaluated.Cohen′s kappa statistics were used to evaluate the consistency of(MALDI-TOF MS)-based rAST method and standard reference BMD.Results:The mass spectrometry of the two methods could successfully identify KP after different incubation time,and the difference was not statistically significant(P>0.05).However,the optimized formic acid extraction method simplified the operation process,and the successful identification rate of the optimized method was higher than that of the conventional scheme under the condition of low bacterial quantity.The MIC of IPM(P<0.01)and TGC(P<0.001)were best measured by(MALDI-TOF MS)-based rAST method at a dilution of 25 times,with a bacterial solution content was 6×10^(6) CFU/mL,and the incubation time was 2 h and 3 h,respectively.Under this condition,the MIC measured by(MALDI-TOF MS)-based rAST and broth microdilution method were similar,IPM 70.83%(85/120),TGC 66.67%(40/60)and essential error(EA)98.33%(118/120),98.33%(59/60),respectively.Conclusion:(MALDI-TOF MS)-based rAST can shorten the reporting time of KP to IPM and TGC,and is a reliable and quantitatively rapid antimicrobial susceptible testing.

关 键 词：肺炎克雷伯菌 MALDI-TOF MS质谱 快速药敏试验 亚胺培南 替加环素 

分 类 号：R372[医药卫生—病原生物学]