Sinzger C, Knapp J, Plachter B, Schmidt K, Jahn G
Abt. für Med. Virologie, Hygiene-Institut, Universität Tübingen, Germany.
J Virol Methods. 1997 Jan;63(1-2):103-12. doi: 10.1016/s0166-0934(97)02082-x.
A method for quantitative analysis of the growth properties of human cytomegalovirus (HCMV) in various cell culture systems was developed. Recent HCMV isolates are, in most cases cell associated, causing only limited cytopathic effect. This renders comparative analysis of the biological properties of such isolates difficult. The focus expansion assay described in this study is based on cocultivation of infected fibroblasts with a cell type of choice, relying on cell mediated infectivity. The extent of replication of a given isolate in cell culture is quantified by determining the size of resulting infectious foci. Analysis of various clinical isolates and laboratory strains indicated that this assay is a reliable and valid method to define growth properties of HCMV in cell culture. Remarkable differences in the cytopathogenicity of these isolates in fibroblasts as well as in endothelial cells were found. The assay will be useful in studies regarding cell tropism and virulence of recent HCMV isolates and for the quick and easy phenotypic characterization of HCMV deletion mutants.
开发了一种用于定量分析人巨细胞病毒（HCMV）在各种细胞培养系统中生长特性的方法。在大多数情况下，近期的HCMV分离株与细胞相关，仅引起有限的细胞病变效应。这使得对这类分离株的生物学特性进行比较分析变得困难。本研究中描述的焦点扩展试验基于将感染的成纤维细胞与选定的细胞类型共同培养，依赖细胞介导的感染性。通过确定产生的感染灶大小来量化给定分离株在细胞培养中的复制程度。对各种临床分离株和实验室菌株的分析表明，该试验是定义HCMV在细胞培养中生长特性的可靠且有效的方法。发现这些分离株在成纤维细胞以及内皮细胞中的细胞致病性存在显著差异。该试验将有助于研究近期HCMV分离株的细胞嗜性和毒力，以及用于HCMV缺失突变体的快速简便的表型鉴定。
