Easy-to-use imaging-cytometry assay to analyze chitin patterns in yeasts

IF 2.2 4区医学 Q3 MYCOLOGY

Journal de mycologie medicale Pub Date : 2024-06-21 DOI:10.1016/j.mycmed.2024.101493

Delphine Aldebert , Bastien Suarez , François Bettega , Emilie Boucher , Cecile Garnaud , Muriel Cornet

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引用次数: 0

Abstract

Background & Aim

Pathogenic fungi are a major threat to public health, and fungal infections are becoming increasingly common and treatment resistant. Chitin, a component of the fungal cell wall, modifies host immunity and contributes to antifungal resistance. Moreover, chitin content is regulated by chitin synthases and chitinases. However, the specific roles and mechanisms remain unclear. In this study, we developed a cytometric imaging assay to quantify chitin content and identify the distribution of chitin in the yeast cell wall.

Methods

The Candida albicans SC5314 and Nakaseomyces glabratus (ex. C. glabrata) ATCC2001 reference strains, as well as 106 clinical isolates, were used. Chitin content, distribution, and morphological parameters were analysed in 12 yeast species. Moreover, machine learning statistical software was used to evaluate the ability of the cytometric imaging assay to predict yeast species using the values obtained for these parameters.

Results

Our imaging-cytometry assay was repeatable, reproducible, and sensitive to variations in chitin content in C. albicans mutants or after antifungal stimulation. The evaluated parameters classified the yeast species into the correct clade with an accuracy of 85 %.

Conclusion

Our findings demonstrate that this easy-to-use assay is an effective tool for the exploration of chitin content in yeast species.

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简便易用的成像-计数测定法，用于分析酵母菌中的几丁质模式。

背景与目的：致病真菌是公共卫生的主要威胁，真菌感染越来越常见，耐药性也越来越强。几丁质是真菌细胞壁的一种成分，它能改变宿主的免疫力并导致抗真菌性。此外，几丁质含量受几丁质合成酶和几丁质酶的调节。然而，其具体作用和机制仍不清楚。在本研究中，我们开发了一种细胞成像检测方法，用于量化几丁质含量并确定几丁质在酵母细胞壁中的分布：方法：使用白念珠菌 SC5314 和 Nakaseomyces glabratus (ex. C. glabrata) ATCC2001 参考菌株以及 106 株临床分离株。分析了 12 种酵母菌的几丁质含量、分布和形态参数。此外，还使用机器学习统计软件评估了细胞成像测定法利用这些参数值预测酵母菌种类的能力：结果：我们的成像-细胞计量测定法具有可重复性和再现性，而且对白茨酵母突变体或抗真菌刺激后几丁质含量的变化很敏感。所评估的参数将酵母菌物种归入正确支系的准确率为 85%：我们的研究结果表明，这种易于使用的检测方法是检测酵母菌几丁质含量的有效工具。

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 de mycologie medicale 医学-真菌学

CiteScore

5.10

自引率

2.80%

发文量

审稿时长

6-12 weeks

期刊介绍： The Journal de Mycologie Medicale / Journal of Medical Mycology (JMM) publishes in English works dealing with human and animal mycology. The subjects treated are focused in particular on clinical, diagnostic, epidemiological, immunological, medical, pathological, preventive or therapeutic aspects of mycoses. Also covered are basic aspects linked primarily with morphology (electronic and photonic microscopy), physiology, biochemistry, cellular and molecular biology, immunochemistry, genetics, taxonomy or phylogeny of pathogenic or opportunistic fungi and actinomycetes in humans or animals. Studies of natural products showing inhibitory activity against pathogenic fungi cannot be considered without chemical characterization and identification of the compounds responsible for the inhibitory activity. JMM publishes (guest) editorials, original articles, reviews (and minireviews), case reports, technical notes, letters to the editor and information. Only clinical cases with real originality (new species, new clinical present action, new geographical localization, etc.), and fully documented (identification methods, results, etc.), will be considered. Under no circumstances does the journal guarantee publication before the editorial board makes its final decision. The journal is indexed in the main international databases and is accessible worldwide through the ScienceDirect and ClinicalKey platforms.

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