Olivo P D
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Clin Microbiol Rev. 1996 Jul;9(3):321-34. doi: 10.1128/CMR.9.3.321.
Rapid diagnostic assays based on direct detection of viral antigen or nucleic acid are being used with increasing frequency in clinical virology laboratories. Virus culture, however, remains the only way to detect infectious virus and to analyze clinically relevant viral phenotypes, such as drug resistance. Growth of viruses in cell culture is labor intensive and time-consuming and requires the use of many different cell lines. Transgenic technology, together with increasing knowledge of the molecular pathways of virus replication, offers the possibility of using genetically modified cell lines to improve virus growth in cell culture and to facilitate detection of virus-infected cells. Genetically modifying cells so that they express a reporter gene only after infection with a specific virus can allow the detection of infectious virus by rapid and simple enzyme assays such as beta-galactosidase assays without the need for antibodies. Although transgenic cells have recently been successfully used for herpes simplex virus detection, much more work needs to be done to adapt this technology to other human viral pathogens such as cytomegalovirus and respiratory viruses. This review offers some strategies for applying this technology to a wide spectrum of animal viruses.
基于直接检测病毒抗原或核酸的快速诊断检测方法在临床病毒学实验室中的使用频率越来越高。然而,病毒培养仍然是检测感染性病毒以及分析临床相关病毒表型(如耐药性)的唯一方法。在细胞培养中培养病毒劳动强度大、耗时且需要使用许多不同的细胞系。转基因技术,加上对病毒复制分子途径的了解不断增加,为利用基因改造的细胞系改善病毒在细胞培养中的生长以及促进病毒感染细胞的检测提供了可能性。对细胞进行基因改造,使其仅在感染特定病毒后才表达报告基因,这样就可以通过快速简单的酶检测(如β-半乳糖苷酶检测)来检测感染性病毒,而无需使用抗体。尽管转基因细胞最近已成功用于单纯疱疹病毒检测,但要使该技术适用于其他人类病毒病原体(如巨细胞病毒和呼吸道病毒),还需要做更多工作。本综述提供了一些将该技术应用于多种动物病毒的策略。