Pathogen and Microbiome Institute, Northern Arizona University, Flagstaff, Arizona.
Curr Protoc Microbiol. 2020 Sep;58(1):e113. doi: 10.1002/cpmc.113.
Coccidioidomycosis ("Valley fever") is caused by Coccidioides immitis and C. posadasii. These fungi are thermally dimorphic, cycling between mycelia and arthroconidia in the environment and converting into spherules and endospores within a host. Coccidioides can cause a broad spectrum of disease that can be difficult to treat. There has been a steady increase in disease, with an estimated 350,000 new infections per year in the United States. With the increase in disease and difficulty in treatment, there is an unmet need to increase research in basic biology and identify new treatments, diagnostics, and vaccine candidates. Here, we describe protocols required in any Coccidioides laboratory, such as growing, harvesting, and storing the different stages of this dimorphic fungal pathogen. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Growth and harvest of liquid mycelia cultures for extractions Alternate Protocol 1: Large-volume growth and harvest of liquid mycelia cultures Basic Protocol 2: Mycelial growth on solid medium Alternate Protocol 2: Maintaining mycelial growth on solid medium Basic Protocol 3: Harvesting and quantification of arthroconidia Alternate Protocol 3: Long-term storage of arthroconidia Basic Protocol 4: Parasitic spherule growth and harvest Alternate Protocol 4: Obtaining endospores from spherules Basic Protocol 5: Intranasal infection of murine models.
球孢子菌病(“山谷热”)是由粗球孢子菌和波氏球孢子菌引起的。这些真菌具有热二态性,在环境中循环为菌丝和分生孢子,在宿主内转化为球体和内生孢子。球孢子菌可引起广泛的疾病,治疗难度大。该病的发病率稳步上升,据估计美国每年有 35 万人新感染。随着疾病的增加和治疗的困难,人们迫切需要增加基础生物学研究,以确定新的治疗方法、诊断方法和疫苗候选物。在这里,我们描述了任何球孢子菌实验室都需要的方案,例如生长、收获和储存这种二态真菌病原体的不同阶段。© 2020 威利父子公司。基本方案 1:提取用液体菌丝培养物的生长和收获备选方案 1:液体菌丝培养物的大量生长和收获基本方案 2:固体培养基上的菌丝生长备选方案 2:固体培养基上的菌丝生长的维持基本方案 3:分生孢子的收获和定量备选方案 3:分生孢子的长期储存基本方案 4:寄生球体的生长和收获备选方案 4:从球体中获得内生孢子基本方案 5:鼠模型的鼻腔内感染