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基质辅助激光解吸电离飞行时间质谱法:一种检测临床分离株对两性霉素B敏感性的快速方法。

MALDI-TOF MS: A Quick Method to Detect the Susceptibility of spp. Clinical Isolates to Amphotericin B.

作者信息

Grizante Barião Patrícia Helena, Cayún Yasna, Sepúlveda Marcela, Tonani Ludmilla, Gonçalves de Almeida Otavio Guilherme, Cornejo Pablo, Dias Nathalia, Santos Cledir, von Zeska Kress Marcia Regina

机构信息

Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão, Ribeirão Preto 14040-903, SP, Brazil.

Programa de Doctorado en Ciencias Mención Biología Celular y Molecular Aplicada, Universidad de La Frontera, Temuco 4811-230, Chile.

出版信息

Microorganisms. 2023 Jul 18;11(7):1834. doi: 10.3390/microorganisms11071834.

Abstract

Disseminated fusariosis is treated with amphotericin B and voriconazole. To determine adequate therapy, the minimal inhibitory concentration (MIC) is used. However, MIC analysis is based on visual observation and requires a long period of fungal incubation. The measure of the minimal profile change concentration (MPCC) using MALDI-TOF MS is a quick spectral method that has presented good results in determining the antimicrobial resistance of yeasts. However, there is a lack of information on filamentous fungi. In the present work, 13 spp. clinical isolates and two reference strains were used. MIC was obtained according to the M38-A2 protocol of the Clinical Laboratory Standards Institute, while MPPC was obtained following the initial steps of the M38-A2 protocol. Both Biotyper and the Rstudio environment were used to analyze mass spectra. For some fungal strains, the data obtained from the software MALDI Biotyper Compass 4.1 led to fuzzy heatmaps resulting in difficult interpretation, while heatmaps obtained using Rstudio tools generated better MPCC resolutions. Herein, 86.6% of the AMB MPCC values were highly correlated with the gold-standard AMB MIC. MALDI-TOF MS is a prominent tool used to determine MPCCs quicker, cost-effectively, and more accurately for spp. strains. However, better statistical analyses could help measure the technique's limit detection.

摘要

播散性镰刀菌病采用两性霉素B和伏立康唑治疗。为确定适当的治疗方法,使用最低抑菌浓度(MIC)。然而,MIC分析基于肉眼观察,且需要较长时间的真菌培养。使用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)测定最低谱图变化浓度(MPCC)是一种快速光谱方法,在确定酵母的抗菌药物耐药性方面已取得良好结果。然而,关于丝状真菌的信息较少。在本研究中,使用了13种临床分离株和两种参考菌株。根据临床实验室标准协会的M38-A2方案获得MIC,而MPPC则按照M38-A2方案的初始步骤获得。使用Biotyper和Rstudio环境分析质谱。对于一些真菌菌株,从软件MALDI Biotyper Compass 4.1获得的数据导致热图模糊,难以解释,而使用Rstudio工具获得的热图产生了更好的MPCC分辨率。在此,86.6%的两性霉素B MPCC值与金标准两性霉素B MIC高度相关。MALDI-TOF MS是一种用于更快、更经济有效且更准确地测定菌株MPCC的重要工具。然而,更好的统计分析有助于衡量该技术的检测限。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f05f/10383446/44b745a91d11/microorganisms-11-01834-g001.jpg

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