MICy：一种快速测定最小抑菌浓度的新型流式细胞术方法。

MICy: a Novel Flow Cytometric Method for Rapid Determination of Minimal Inhibitory Concentration.

机构信息

Department of Anaesthesiology and Intensive Therapy, Semmelweis Universitygrid.11804.3c, Budapest, Hungary.

Department of Physiology, Semmelweis Universitygrid.11804.3c, Budapest, Hungary.

出版信息

Microbiol Spectr. 2021 Dec 22;9(3):e0090121. doi: 10.1128/spectrum.00901-21. Epub 2021 Dec 8.

DOI:10.1128/spectrum.00901-21

PMID:34878335

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8653823/

Abstract

Early initiated adequate antibiotic treatment is essential in intensive care. Shortening the length of antibiotic susceptibility testing (AST) can accelerate clinical decision-making. Our objective was to develop a simple flow cytometry (FC)-based AST that produces reliable results within a few hours. We developed a FC-based AST protocol (MICy) and tested it on six different bacteria strains (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis) in Mueller-Hinton and Luria-Bertani broth. We monitored the bacterial growth by FC to define the optimal time of AST. All bacteria were tested against 12 antibiotics and the MIC values were compared to microdilution used as reference method. McNemar and Fleiss' kappa inter-observer tests were performed to analyze the bias between the two methods. Susceptibility profiles of the two methods were also compared. We found that FC is able to detect the bacterial growth after 4-h incubation. The point-by-point comparison of MICy and microdilution resulted in exact match above 87% (2642/3024) of all measurements. The MIC values obtained by MICy and microdilution agreed over 80% (173/216) within ±1 dilution range that gives a substantial inter-observer agreement with weighted Fleiss' kappa. By using the EUCAST clinical breakpoints, we defined susceptibility profiles of MICy that were identical to microdilution in more than 92% (197/213) of the decisions. MICy resulted 8.7% major and 3.2% very major discrepancies. MICy is a new, simple FC-based AST method that produces susceptibility profile with low failure rate a workday earlier than the microdilution method. MICy is a new, simple and rapid flow cytometry based antibiotic susceptibility testing (AST) method that produces susceptibility profile a workday earlier than the microdilution method or other classical phenotypic AST methods. Shortening the length of AST can accelerate clinical decision-making as targeted antibiotic treatment improves clinical outcomes and reduces mortality, duration of artificial ventilation, and length of stay in intensive care unit. It can also reduce nursing time and costs and the spreading of antibiotic resistance. In this study, we present the workflow and methodology of MICy and compare the results produced by MICy to microdilution step by step.

摘要

早期开始适当的抗生素治疗是重症监护的关键。缩短抗生素药敏试验（AST）的时间可以加速临床决策。我们的目标是开发一种简单的基于流式细胞术（FC）的 AST，能够在数小时内产生可靠的结果。我们开发了一种基于 FC 的 AST 方案（MICy），并在 Mueller-Hinton 和 Luria-Bertani 肉汤中对六种不同的细菌菌株（大肠杆菌、肺炎克雷伯菌、铜绿假单胞菌、金黄色葡萄球菌、化脓性链球菌、粪肠球菌）进行了测试。我们通过 FC 监测细菌生长，以确定 AST 的最佳时间。所有细菌均进行了 12 种抗生素的测试，MIC 值与作为参考方法的微量稀释法进行比较。采用 McNemar 和 Fleiss' kappa 观察者间检验分析两种方法之间的偏差。还比较了两种方法的药敏谱。我们发现 FC 能够在孵育 4 小时后检测到细菌生长。MICy 和微量稀释法的点对点点比较产生了超过 87%（2642/3024）的所有测量结果完全一致。MICy 和微量稀释法获得的 MIC 值在±1 稀释范围内一致，超过 80%（173/216），具有实质性的观察者间一致性，加权 Fleiss' kappa 值为 0.76。使用 EUCAST 临床折点，我们定义了 MICy 的药敏谱，在超过 92%（197/213）的决策中与微量稀释法相同。MICy 导致 8.7%的主要和 3.2%的非常主要差异。MICy 是一种新的、简单的基于 FC 的 AST 方法，与微量稀释法相比，能在工作日提前一天产生失败率较低的药敏谱。MICy 是一种新的、简单和快速的基于流式细胞术的抗生素药敏试验（AST）方法，与微量稀释法或其他经典表型 AST 方法相比，能在工作日提前一天产生药敏谱。缩短 AST 的时间可以加速临床决策，因为靶向抗生素治疗可以改善临床结果，降低死亡率、人工通气时间和重症监护病房的住院时间。它还可以减少护理时间和成本，以及抗生素耐药性的传播。在这项研究中，我们介绍了 MICy 的工作流程和方法学，并逐步比较了 MICy 和微量稀释法产生的结果。