Infection and Propagation of Astrovirus VA1 in Cell Culture.

Current Protocols in Microbiology Pub Date : 2019-02-01 Epub Date: 2018-11-16 DOI:10.1002/cpmc.73
Andrew B Janowski, David Wang
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Abstract

Astrovirus VA1/HMO-C (VA1) is the representative genotype of mamastrovirus 9, a species of the single-stranded, positive-sense RNA viral family, Astroviridae. Astroviruses have been traditionally considered pathogens of the gastrointestinal tract but they have been recently associated with neurological diseases in humans, cattle, mink, sheep, and pigs. VA1 is the astrovirus genotype most commonly identified from human cases of meningoencephalitis and has been recently propagated in cell culture. VA1 can now be used as a model system to study pathogenesis of the neurological diseases associated with astrovirus infection. In this article, we describe two fundamental assays to quantify replication and propagation of VA1, a quantitative reverse transcription-PCR (qRT-PCR) to measure viral RNA and a 50% tissue culture infectious dose (TCID50 ) assay to measure infectious viral particles. © 2018 by John Wiley & Sons, Inc.

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天体病毒 VA1 在细胞培养中的感染和繁殖。
天体病毒 VA1/HMO-C(VA1)是天体病毒科(Astroviridae)单链正义 RNA 病毒属(mamastrovirus 9)的代表基因型。天体病毒传统上被认为是胃肠道的病原体,但近来却与人类、牛、水貂、绵羊和猪的神经系统疾病有关。VA1 是最常从人类脑膜脑炎病例中发现的星状病毒基因型,最近已在细胞培养中繁殖。现在,VA1 可用作研究与星状病毒感染相关的神经系统疾病发病机制的模型系统。在这篇文章中,我们介绍了量化VA1复制和繁殖的两种基本检测方法,一种是定量反转录-PCR(qRT-PCR)检测病毒RNA,另一种是50%组织培养感染剂量(TCID50)检测感染性病毒颗粒。© 2018 by John Wiley & Sons, Inc.
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Current Protocols in Microbiology
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.
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