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用于隐孢子虫感染研究及利用流式细胞术、qPCR和组织病理学对寄生虫负荷进行定量的小鼠模型。

Mouse Models for Use in Cryptosporidium Infection Studies and Quantification of Parasite Burden Using Flow Cytometry, qPCR, and Histopathology.

作者信息

Sonzogni-Desautels Karine, Mead Jan R, Ndao Momar

机构信息

National Reference Centre for Parasitology, Research Institute of the McGill University Health Centre, Montreal, QC, Canada.

Infectious Diseases and Immunity in Global Health Program, Research Institute of the McGill University Health Centre, Montreal, QC, Canada.

出版信息

Methods Mol Biol. 2020;2052:229-251. doi: 10.1007/978-1-4939-9748-0_14.

Abstract

Cryptosporidiosis threatens life of young children in developing countries and newborn calves around the world. No vaccine or therapy can prevent or cure this diarrhea-inducing enteric disease caused by Cryptosporidium spp. protozoan parasites. There is an essential need to discover new therapeutic drugs efficient in reducing parasite burden in infected individuals. Research therefore relies on reliable small animal models of cryptosporidiosis. Here, we present excellent mouse models which can efficiently mimic pathogenesis of human and bovine cryptosporidiosis. We also describe methods to purify C. parvum oocysts from stool and intestine of infected mice to facilitate oocyst quantification. Moreover, we present protocols using flow cytometry, quantitative polymerase chain reaction, and histopathology to accurately quantify parasite burden in stool or intestine samples.

摘要

隐孢子虫病威胁着发展中国家幼儿以及世界各地新生小牛的生命。尚无疫苗或疗法能够预防或治愈这种由隐孢子虫属原生动物寄生虫引起的导致腹泻的肠道疾病。迫切需要发现能够有效减轻感染者体内寄生虫负担的新型治疗药物。因此,研究依赖于可靠的隐孢子虫病小动物模型。在此,我们展示了能够有效模拟人类和牛隐孢子虫病发病机制的优秀小鼠模型。我们还描述了从感染小鼠的粪便和肠道中纯化微小隐孢子虫卵囊以促进卵囊定量的方法。此外,我们介绍了使用流式细胞术、定量聚合酶链反应和组织病理学来准确量化粪便或肠道样本中寄生虫负担的方案。

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