Cannon M J
Division of Immunology, National Institute for Medical Research, London, U.K.
J Virol Methods. 1987 Jul;16(4):293-301. doi: 10.1016/0166-0934(87)90014-0.
A rapid microplaque technique was developed for the detection of infectious respiratory syncytial virus (RSV) in the lungs of infected mice. Infected lung homogenates were titrated on microwell HEp-2 monolayers and incubated for 24 or 48 h. The microwells were then fixed with 4% formaldehyde in saline, or methanol containing 0.5% hydrogen peroxide. 24-h single cell infectious foci and 48-h microplaques were detected by an indirect immunoperoxidase (IIP) assay using monoclonal antibodies specific for RSV envelope glycoproteins as the first layer. This method can be used for the quantification of lung RSV in large numbers of samples much more rapidly and economically than conventional plaque assay techniques. In addition, the use of the IIP assay renders the system specific for RSV.
开发了一种快速微斑技术，用于检测感染小鼠肺部的传染性呼吸道合胞病毒（RSV）。将感染的肺匀浆在微孔HEp-2单层上滴定，并孵育24或48小时。然后用盐水中的4%甲醛或含0.5%过氧化氢的甲醇固定微孔。使用针对RSV包膜糖蛋白的单克隆抗体作为第一层，通过间接免疫过氧化物酶（IIP）测定法检测24小时的单细胞感染灶和48小时的微斑。与传统的噬斑测定技术相比，该方法可更快速、经济地用于大量样品中肺RSV的定量。此外，IIP测定法的使用使该系统对RSV具有特异性。
