A rapid and simple MALDI-TOF MS lipid profiling method for differentiating Mycobacterium ulcerans from Mycobacterium marinum.

IF 6.1 2区 医学 Q1 MICROBIOLOGY
Journal of Clinical Microbiology Pub Date : 2025-03-12 Epub Date: 2025-01-27 DOI:10.1128/jcm.01400-24
Takeshi Komine, Hanako Fukano, Mitsunori Yoshida, Yuji Miyamoto, Makoto Nakaya, Azumi Fujinaga, Kohei Doke, Yoshihiko Hoshino
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引用次数: 0

Abstract

Mycobacterium ulcerans, a slow-growing nontuberculous mycobacterium, causes Buruli ulcer, a neglected tropical disease. Distinguishing M. ulcerans from related species, including Mycobacterium marinum, poses challenges with respect to making accurate identifications. In this study, we developed a rapid and simple identification method based on mycobacterial lipid profiles and used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to analyze the lipid profiles of M. ulcerans (n = 35) and M. marinum (n = 19) isolates. Bacterial colonies pre-cultured on 2% Ogawa egg slants for 2 months were collected, and total lipids were extracted using an MBT Lipid Xtract kit. Spectra were obtained in negative ion mode using a MALDI Biotyper Sirius system, with ClinProTools v3.0 being used to analyze the spectra based on the application of three algorithms (genetic algorithm [GA], supervised neural network [SNN], and quick classifier [QC)]). Cross-validation was performed using a 20% leave-out set randomly selected from the samples. Models generated using GA, SNN, and QC showed cross-validation values of 100%, 100%, and 97.9%, respectively, and all algorithms achieved 100% recognition capability values. Our findings indicate that MALDI-TOF analysis of lipid profiles can accurately differentiate two mycobacterial species (M. ulcerans and M. marinum) that are difficult to distinguish using conventional protein-targeting methods.IMPORTANCEBuruli ulcer, caused by Mycobacterium ulcerans, is a neglected tropical disease. However, distinguishing M. ulcerans from related species, including Mycobacterium marinum, presents certain challenges. In this study, we demonstrate the utility of a rapid yet simple method for differentiating isolates of these mycobacteria based on their lipid profiles using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. This new approach can accurately identify species that are otherwise difficult to distinguish using conventional techniques. This represents a significant diagnostic advance for clinical laboratories, in that it enables a more rapid and precise identification, thereby leading to earlier treatment initiation and more appropriate treatment regimens for infections caused by these bacteria.

快速简便的MALDI-TOF质谱法鉴别溃疡分枝杆菌与海洋分枝杆菌。
溃疡分枝杆菌是一种生长缓慢的非结核分枝杆菌,可引起布鲁里溃疡,这是一种被忽视的热带疾病。将溃疡分枝杆菌与包括海洋分枝杆菌在内的相关物种区分开来,对准确鉴定提出了挑战。在这项研究中,我们建立了一种基于分枝杆菌脂质谱的快速、简单的鉴定方法,并使用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)分析了溃疡分枝杆菌(35株)和海洋分枝杆菌(19株)的脂质谱。收集在2%小川鸡蛋斜面上预培养2个月的菌落,用MBT脂质提取试剂盒提取总脂质。使用MALDI Biotyper Sirius系统在负离子模式下获得光谱,使用ClinProTools v3.0进行光谱分析,采用遗传算法[GA]、监督神经网络[SNN]和快速分类器[QC]三种算法。使用从样本中随机选择的20%遗漏集进行交叉验证。使用GA、SNN和QC生成的模型交叉验证值分别为100%、100%和97.9%,所有算法均达到100%的识别能力值。我们的研究结果表明,MALDI-TOF分析脂质谱可以准确区分两种分枝杆菌(溃疡分枝杆菌和海洋分枝杆菌),这两种分枝杆菌很难用传统的蛋白质靶向方法区分。由溃疡分枝杆菌引起的布鲁里溃疡是一种被忽视的热带病。然而,区分溃疡分枝杆菌与相关物种,包括海洋分枝杆菌,提出了一定的挑战。在这项研究中,我们利用基质辅助激光解吸/电离飞行时间质谱法,证明了一种快速而简单的方法,可以根据这些分枝杆菌的脂质谱来区分它们。这种新方法可以准确地识别出用传统技术难以区分的物种。这代表了临床实验室在诊断方面的重大进步,因为它能够更快速和准确地识别,从而导致更早地开始治疗,并为这些细菌引起的感染提供更适当的治疗方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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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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