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在 WASPLab 上短时间孵育后,通过 MALDI-TOF/MS 快速鉴定和抗菌药敏纸片扩散法对阳性血培养物进行药敏试验。

Rapid identification by MALDI-TOF/MS and antimicrobial disk diffusion susceptibility testing for positive blood cultures after a short incubation on the WASPLab.

机构信息

Department of Diagnostics, Bacteriology Laboratory, Division of Laboratory Medicine, Geneva University Hospitals, 4 rue Gabrielle-Perret-Gentil, 1205, Geneva, Switzerland.

Department of Diagnostics, Division of Laboratory Medicine, Geneva University Hospitals, Geneva, Switzerland.

出版信息

Eur J Clin Microbiol Infect Dis. 2020 Jun;39(6):1063-1070. doi: 10.1007/s10096-020-03817-8. Epub 2020 Jan 21.

Abstract

The objectives of this study were to define the shortest incubation times on the WASPLab for reliable MALDI-TOF/MS-based species identification and for the preparation of a 0.5 McFarland suspension for antimicrobial disk diffusion susceptibility testing using short subcultures growing on solid culture media inoculated by positive blood cultures spiked with a wide range of pathogens associated with bloodstream infections. The 520 clinical strains (20 × 26 different species) included in this study were obtained from a collection of non-consecutive and non-duplicate pathogens identified at Geneva University Hospitals. After 4 h of incubation on the WASPLab, microorganisms' growth allowed accurate identification of 73% (380/520) (95% CI, 69.1-76.7%) of the strains included in this study. The identification rate increased to 85% (440/520) (95% CI, 81.3-87.5%) after 6-h incubation. When excluding Corynebacterium and Candida spp., the microbial growth was sufficient to permit accurate identification of all tested species (100%, 460/460) (95% CI, 99.2-100%) after 8-h incubation. With the exception of Burkholderia cepacia and Haemophilus influenzae, AST by disk diffusion could be performed for Enterobacterales and non-fermenting Gram-negative bacilli after only 4 h of growth in the WASPLab. The preparation of a 0.5 McFarland suspension for Gram-positive bacteria required incubation times ranging between 3 and 8 h according to the bacterial species. Only Corynebacterium spp. required incubation times as long as 16 h. The WASPLab enables rapid pathogen identification as well as swift comprehensive AST from positive blood cultures that can be implemented without additional costs nor hands-on time by defining optimal time points for image acquisition.

摘要

本研究的目的是确定 WASPLab 上进行可靠 MALDI-TOF/MS 物种鉴定的最短孵育时间,以及用于制备 0.5 McFarland 混悬液的最短孵育时间,该混悬液用于使用固体培养基于短时间培养物进行抗菌药物纸片扩散药敏试验,这些短时间培养物是从日内瓦大学医院鉴定的一系列连续和非重复血培养阳性污染的病原体中接种的。本研究共纳入 520 株临床分离株(20×26 种不同的物种),这些菌株来自日内瓦大学医院收集的非连续和非重复的病原体。在 WASPLab 孵育 4 小时后,微生物的生长允许对本研究中包含的 73%(380/520)(95%CI,69.1-76.7%)的菌株进行准确鉴定。孵育 6 小时后,鉴定率增加至 85%(440/520)(95%CI,81.3-87.5%)。排除棒状杆菌和假丝酵母后,孵育 8 小时后,足够的微生物生长可允许对所有测试的物种(100%,460/460)(95%CI,99.2-100%)进行准确鉴定。除伯克霍尔德菌和流感嗜血杆菌外,在 WASPLab 中培养 4 小时后,肠杆菌科和非发酵革兰氏阴性杆菌可进行纸片扩散药敏试验。革兰氏阳性菌制备 0.5 McFarland 混悬液的孵育时间根据细菌种类在 3 至 8 小时之间。只有棒状杆菌需要长达 16 小时的孵育时间。WASPLab 可以实现快速的病原体鉴定以及快速的全面药敏试验,这可以通过定义最佳的图像采集时间点来实现,而无需额外的成本或人力。

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