市售培养基对肉汤微量稀释法头孢地罗药敏试验的影响。

IF 5.4 2区 医学 Q1 MICROBIOLOGY
Journal of Clinical Microbiology Pub Date : 2025-09-10 Epub Date: 2025-08-20 DOI:10.1128/jcm.00471-25
Boudewijn L M DeJonge, Christine Slover, Sean T Nguyen, Christopher Longshaw, Miki Takemura, Naomi Anan, Hidenori Yamashiro, Yoshinori Yamano
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引用次数: 0

摘要

测定头孢地洛尔MIC的参考方法是用缺铁阳离子调整穆勒-辛顿肉汤(ID-CAMHB)进行肉汤微量稀释(BMD)。然而,实验室报告了不同的MIC结果和确定MIC终点的困难。为了研究这一点,通过检查培养基中铁耗尽的螯合时间,并使用4种MHB源(BD-BBL、BD-Difco、Oxoid和Merck)和85种革兰氏阴性分离物(大肠杆菌、肺炎克雷伯菌、铜绿假单胞菌和鲍曼不动杆菌)测试再现性,对BMD方法进行了重新评估,其中63种先前在体内进行了评估。将螯合时间延长至6小时,所有介质的铁还原效果最佳,≤0.03µg/mL。每种培养基源的MIC重现性都很高,且因物种而异(在模态MIC值的一次稀释内,MIC值为93.3%-99.0%);然而,不同来源的ID-CAMHB对分离株产生(可重复的)不同的模态MIC值。BD-BBL和BD-Difco的ID-CAMHB相关性最好,96.5%的分离株在培养基之间的一次稀释内显示模态MIC值。这两种培养基产生的MIC值最能预测体内疗效。当观察到拖尾时,在所有介质中都可以看到,并应用了精确的读数指南来提高确定MIC终点的可重复性。6-h螯合和精炼的阅读指南,拖尾现在纳入临床和实验室标准研究所M100文件。采用BMD法测定头孢地罗药敏性时,应考虑介质的影响,基于这些结果,推荐使用BD-BBL或BD-Difco制备的ID-CAMHB。头孢地罗的参考药敏试验方法为2016年临床与实验室标准研究所批准的肉汤微量稀释法,使用缺铁阳离子调整的Mueller-Hinton肉汤。一些复制这种方法的手动设备已经进入市场,但这些设备的结果不一致。本研究将穆勒-辛顿肉汤的来源确定为头孢德罗col MIC测定的一个变量,并详细描述了缺铁阳离子调整穆勒-辛顿肉汤的制备方法,并修订了读数指南,以提高头孢德罗col MIC测定对大肠杆菌、肺炎克雷伯菌、铜绿假单胞菌和鲍曼不动态杆菌的重复性。本研究提出的建议可提高头孢地罗微稀释药敏试验的重复性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The impact of commercially available media on cefiderocol susceptibility testing by broth microdilution method.

The reference method for determining cefiderocol MIC is broth microdilution (BMD) using iron-depleted cation-adjusted Mueller-Hinton broth (ID-CAMHB). However, laboratories have reported variable MIC findings and difficulties in determining MIC endpoints. To investigate this, the BMD method was re-evaluated by examining the chelation time for iron depletion of the media and by testing reproducibility using four MHB sources (BD-BBL, BD-Difco, Oxoid, and Merck) and 85 Gram-negative isolates (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii), of which 63 were previously evaluated in vivo. Extending chelation time to 6 h provided optimal iron reduction to ≤0.03 µg/mL for all media. MIC reproducibility was high for each medium source and varied by species (93.3%-99.0% of MIC values within one dilution of modal MIC values); however, different sources of ID-CAMHB produced (reproducible) different modal MIC values for isolates. ID-CAMHB from BD-BBL and BD-Difco showed the best correlation with 96.5% of isolates showing modal MIC values within one dilution between media. MIC values generated with these two media were the most predictive of efficacy in vivo. When trailing was observed, it was seen across all media, and refined reading guidelines were applied to enhance reproducibility of determining the MIC endpoint. The 6-h chelation and the refined reading guidelines for trailing are now incorporated into the Clinical and Laboratory Standards Institute M100 document. When using the BMD method to determine cefiderocol susceptibility, the impact of media should be considered, and based on these results, ID-CAMHB prepared with BD-BBL or BD-Difco is recommended.IMPORTANCEThe reference antibiotic susceptibility testing method for cefiderocol is the broth microdilution method approved by the Clinical and Laboratory Standards Institute in 2016, using iron-depleted cation-adjusted Mueller-Hinton broth. A few manual devices have come to the market replicating this method, but inconsistent results with these devices have been reported. The current study identified the source of Mueller-Hinton broth as a variable in cefiderocol MIC determinations and provides detailed description of the preparation of iron-depleted cation-adjusted Mueller-Hinton broth, and revised reading guidance to improve reproducibility of cefiderocol MIC determinations for Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii. The recommendations made in this study should enhance the reproducibility of cefiderocol broth microdilution susceptibility testing.

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来源期刊
Journal of Clinical Microbiology
Journal of Clinical Microbiology 医学-微生物学
CiteScore
17.10
自引率
4.30%
发文量
347
审稿时长
3 months
期刊介绍: The Journal of Clinical Microbiology® disseminates the latest research concerning the laboratory diagnosis of human and animal infections, along with the laboratory's role in epidemiology and the management of infectious diseases.
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