Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.
Institute of Virology, Ulm University Medical Center, Ulm, Germany.
Methods Mol Biol. 2021;2244:19-38. doi: 10.1007/978-1-0716-1111-1_2.
Human cytomegalovirus is routinely isolated by inoculating fibroblast cultures with clinical specimens suspected of harboring HCMV and then monitoring the cultures for cytopathic effects characteristic of this virus. Initially, such clinical isolates are usually strictly cell-associated, but continued propagation in cell culture increases the capacity of an HCMV isolate to release cell-free infectious progeny. Once cell-free infection is possible, genetically homogenous virus strains can be purified by limiting dilution infections. HCMV strains can differ greatly with regard to the titers that can be achieved, the tropism for certain cell types, and the degree to which nonessential genes have been lost during propagation. As there is no ideal HCMV strain for all purposes, the choice of the most appropriate strain depends on the requirements of the particular experiment or project. In this chapter, we provide information that can serve as a basis for deciding which strain may be the most appropriate for a given experiment.
人巨细胞病毒通常通过将临床标本接种到成纤维细胞培养物中进行分离,这些标本被怀疑携带 HCMV,然后监测培养物中是否有该病毒特有的细胞病变效应。最初,这种临床分离株通常是严格细胞相关的,但在细胞培养中继续繁殖会增加 HCMV 分离株释放无细胞感染性后代的能力。一旦可以进行无细胞感染,就可以通过限制稀释感染来纯化遗传同质的病毒株。HCMV 株在可达到的滴度、对某些细胞类型的嗜性以及在繁殖过程中非必需基因丢失的程度方面差异很大。由于没有一种 HCMV 株适合所有目的,因此选择最合适的株取决于特定实验或项目的要求。在本章中,我们提供了一些信息,可以作为决定哪种株最适合给定实验的基础。