研究白色念珠菌发病机制的简单线虫感染模型

Current Protocols in Microbiology Pub Date : 2020-09-25 DOI:10.1002/cpmc.114

Grace H. Kim, Sierra Rosiana, Natalia V. Kirienko, Rebecca S. Shapiro

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引用次数: 7

摘要

白色念珠菌是一种机会性真菌病原体，是研究真菌发病机制的模式生物。它作为一种无害的共生生物和健康人体微生物组的成员存在，但可引起危及生命的粘膜和全身感染。研究白色念珠菌感染及其发病机制的模型宿主是秀丽隐杆线虫。秀丽隐杆线虫经常被用作研究微生物-宿主相互作用的模型宿主，因为它可以被许多人类病原体感染，而且秀丽隐杆线虫和哺乳动物的肠道细胞在形态上也有密切的相似之处，白色念珠菌感染也可能发生。本文概述了利用秀丽隐杆线虫作为宿主研究白色念珠菌感染的详细方法，包括秀丽隐杆线虫卵制备方案和基于琼脂的秀丽隐杆线虫杀死方案，以监测真菌毒力。这些方案还可以用于研究白色念珠菌的基因突变，以进一步了解与白色念珠菌发病和毒力有关的基因以及宿主-微生物相互作用的机制。©2020 Wiley期刊有限公司基本方案1:秀丽隐杆线虫卵的制备支持方案1:冷冻和恢复秀丽隐杆线虫支持方案2:制作超级食品琼脂和OP50板基本方案2:秀丽隐杆线虫/白色念珠菌琼脂杀灭试验支持方案3:构建虫挑

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A Simple Nematode Infection Model for Studying Candida albicans Pathogenesis

Candida albicans is an opportunistic fungal pathogen and a model organism to study fungal pathogenesis. It exists as a harmless commensal organism and member of the healthy human microbiome, but can cause life-threatening mucosal and systemic infections. A model host to study C. albicans infection and pathogenesis is the nematode Caenorhabditis elegans. C. elegans is frequently used as a model host to study microbial-host interactions because it can be infected by many human pathogens and there are also close morphological resemblances between the intestinal cells of C. elegans and mammals, where C. albicans infections can occur. This article outlines a detailed methodology for exploiting C. elegans as a host to study C. albicans infection, including a C. elegans egg preparation protocol and an agar-based C. elegans killing protocol to monitor fungal virulence. These protocols can additionally be used to study C. albicans genetic mutants in order to further our understanding of the genes involved in pathogenesis and virulence in C. albicans and the mechanisms of host-microbe interactions. © 2020 Wiley Periodicals LLC.

Basic Protocol 1: Preparation of Caenorhabditis elegans eggs

Support Protocol 1: Freezing and recovering Caenorhabditis elegans

Support Protocol 2: Making superfood agar and OP50 plates

Basic Protocol 2: Caenorhabditis elegans/Candida albicans agar killing assay

Support Protocol 3: Constructing a worm pick

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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.