Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA, USA.
Methods Mol Biol. 2020;2123:281-294. doi: 10.1007/978-1-0716-0346-8_20.
Influenza virus isolation is a procedure to obtain a live and infectious virus that can be used for antigenic characterization, pathogenesis investigation, vaccine production, and so on. Embryonated chicken egg inoculation is traditionally considered the "gold standard" method for influenza virus isolation and propagation. However, many primary cells and continuous cell lines have also been examined or developed for influenza virus isolation and replication. Specifically, influenza A virus in swine (IAV-S) isolation and propagation has been attempted and compared in embryonated chicken eggs, some primary porcine cells, and a number of continuous cell lines. Currently, Madin-Darby canine kidney (MDCK) cells remain the most commonly used cell line for the isolation, propagation, and titration of IAV-S. Virus isolation in embryonated chicken eggs or in different cell lines offers alternative approaches when IAV-S isolation in MDCK cells is unsuccessful. Optimal specimens for IAV-S isolation includes nasal swabs, nasopharyngeal swabs, oral fluids, bronchoalveolar lavage, lung tissues, and so on. In this chapter, we describe the procedures of sample processing, IAV-S isolation in MDCK cells and in embryonated chicken eggs, as well as the methods used for confirming the virus isolation results.
流感病毒分离是一种获取活的、有感染性的病毒的方法，可用于抗原特征分析、发病机制研究、疫苗生产等。鸡胚接种传统上被认为是流感病毒分离和繁殖的“金标准”方法。然而，许多原代细胞和连续细胞系也被用于流感病毒的分离和复制。具体来说，已经尝试并比较了猪流感病毒（IAV-S）在鸡胚、一些原代猪细胞和一些连续细胞系中的分离和繁殖。目前，Madin-Darby 犬肾（MDCK）细胞仍然是用于 IAV-S 分离、繁殖和滴定的最常用的细胞系。当 MDCK 细胞中无法分离出 IAV-S 时，鸡胚或不同细胞系中的病毒分离为提供了替代方法。IAV-S 分离的最佳标本包括鼻拭子、鼻咽拭子、口腔液、支气管肺泡灌洗、肺组织等。在本章中，我们描述了样品处理、MDCK 细胞和鸡胚中 IAV-S 分离以及用于确认病毒分离结果的方法的步骤。
