新型隐球菌抗真菌耐药性的实验进化

Current Protocols in Microbiology Pub Date : 2020-09-28 DOI:10.1002/cpmc.116

Arianne Bermas, Rebecca S. Shapiro, Jennifer Geddes-McAlister

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引用次数: 3

摘要

新型隐球菌是一种机会性酵母样真菌病原体，已证明对用于治疗隐球菌性脑膜炎的所有主要类别的抗真菌药物具有耐药性。然而，由于抗真菌耐药菌株的进化，与这种真菌疾病作斗争是临床医生面临的一个持续挑战。临床批准的抗真菌药物的有限可用性提高了研究耐药分子机制的紧迫性。通过实验进化研究真菌病原体如何在体外进化为抗真菌药物，为研究抗真菌耐药性机制提供了一种简单而有力的方法。实验进化涉及在实验室条件下微生物种群的连续传代，这样可以发生适应性突变并实时监测。这项技术在研究新生C.真菌耐药机制中起着关键作用，这有助于开发新的策略来对抗耐药性的出现。在这里，我们概述了如何进行夜间培养和如何进行实验进化，我们提出了一种分光光度分析来评估抗真菌耐药性的进化。©2020 Wiley期刊公司基本方案1:新隐球菌的生长和样品制备基本方案2:抗真菌耐药性的实验进化基本方案3:抗真菌耐药性的进化分析基本方案4:甘油原液制备

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Experimental Evolution of Antifungal Resistance in Cryptococcus neoformans

Cryptococcus neoformans, an opportunistic yeast-like fungal pathogen, has demonstrated resistance to all major classes of antifungals used to treat cryptococcal meningitis. However, combatting this fungal disease is an ongoing challenge among clinicians due to the evolution of antifungal-resistant strains. The limited availability of clinically approved antifungals has heightened the urgency to investigate the molecular mechanisms underscoring resistance. Studying how a fungal pathogen evolves to an antifungal drug in vitro using experimental evolution provides a simple, yet powerful approach to study the mechanisms of antifungal resistance. Experimental evolution involves the serial passaging of microbial populations under laboratory conditions, such that adaptive mutations can occur and be monitored in real time. This technique plays a key role in investigating the mechanisms of antifungal resistance in C. neoformans, and this can help in developing novel strategies to combat the emergence of resistance. Here, we outline how to make overnight cultures of C. neoformans and how to perform experimental evolution, and we present a spectrophotometric analysis to evaluate the evolution of antifungal resistance. © 2020 Wiley Periodicals LLC.

Basic Protocol 1: Growth and sample preparation of Cryptococcus neoformans

Basic Protocol 2: Experimental evolution of antifungal resistance

Basic Protocol 3: Analyzing the evolution of antifungal resistance

Basic Protocol 4: Glycerol stock preparation

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来源期刊

Current Protocols in Microbiology Immunology and Microbiology-Parasitology

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期刊介绍： Current Protocols in Microbiology provides detailed, step-by-step instructions for analyzing bacteria, animal and plant viruses, fungi, protozoans and other microbes. It offers updated coverage of emerging technologies and concepts, such as biofilms, quorum sensing and quantitative PCR, as well as proteomic and genomic methods. It is the first comprehensive source of high-quality microbiology protocols that reflects and incorporates the new mandates and capabilities of this robust and rapidly evolving discipline.