Lee Wai-Ming, Chen Yin, Wang Wensheng, Mosser Anne
Biological Mimetics Inc., 124 Byte Drive, Frederick, MD, 21702, USA,
Methods Mol Biol. 2015;1221:71-81. doi: 10.1007/978-1-4939-1571-2_7.
Infectivity is a fundamental property of viral pathogens such as human rhinoviruses (HRVs). This chapter describes two methods for measuring the infectivity of HRV-A and -B serotypes: end point dilution (TCID50) assay and plaque assay. End point dilution assay is a quantal, not quantitative, assay that determines the dilution of the sample at which 50 % of the aliquots have infectious virus. It can be used for all the HRV-A and -B serotypes and related clinical isolates that grow in cell culture and induce cytopathic effect (CPE), degenerative changes in cells that are visible under a microscope. Plaque assay is a quantitative assay that determines the number of infectious units of a virus in a sample. After an infectious unit of virus infects one cell, the infected cell produces progeny viruses that then infect and kill a circle of adjacent cells. This circle of dead cells detaches from the dish and thus leaves a clear hole in a cell monolayer. Plaque assay works only for HeLa-adapted HRV-A and -B serotypes that can make visible plaques on the cell monolayer. Currently the end point dilution assay and plaque assay have not been developed for the newly discovered HRV-C.
传染性是人类鼻病毒(HRV)等病毒病原体的基本特性。本章介绍了两种检测HRV - A和 - B血清型传染性的方法:终点稀释(TCID50)试验和平板试验。终点稀释试验是一种定性而非定量的试验,它确定样品的稀释度,在该稀释度下50%的等分试样含有感染性病毒。它可用于所有能在细胞培养中生长并诱导细胞病变效应(CPE)的HRV - A和 - B血清型及相关临床分离株,细胞病变效应是指在显微镜下可见的细胞退行性变化。平板试验是一种定量试验,用于确定样品中病毒感染单位的数量。病毒的一个感染单位感染一个细胞后,被感染的细胞产生子代病毒,这些子代病毒随后感染并杀死一圈相邻的细胞。这一圈死亡的细胞从培养皿上脱离,从而在细胞单层上留下一个清晰的孔。平板试验仅适用于适应HeLa细胞的HRV - A和 - B血清型,它们能在细胞单层上形成可见的噬斑。目前尚未为新发现的HRV - C开发终点稀释试验和平板试验。