Flow cytometry-based assay for titrating dengue virus
Flow cytometry-based assay for titrating dengue virus
复制标题
DOI：
10.1128/jcm.43.7.3267-3272.2005
复制
发表时间：
2005-07-01
影响因子：
9.4
通讯作者：
de Silva, AM
中科院分区：
文献类型：
--
作者：
Lambeth, CR;White, LJ;de Silva, AM
用于测定登革病毒（DENV）的空斑试验耗时且不适合不空斑的菌株。荧光活化细胞分选（FACS）已被用于检测denv感染的细胞。在这里，我们描述了一种基于facs的测定DENV的方法。我们确定在感染后24小时，FACS检测到的感染细胞数量代表了第一轮感染，因此可以用作接种物中感染颗粒数量的读数。当使用FACS、斑块和终点稀释法比较不同实验室和临床DENV菌株的滴度时，大多数菌株的FACS滴度与斑块或终点稀释法获得的滴度相当。FACS测定法是对菌斑测定法的改进，因为感染期从5 - 7天减少到24小时，该测定法可用于滴定临床分离株，这些分离株通常不会在细胞单层上形成清晰的菌斑。本文描述的基于facs的新方法将促进登革热的实验室研究。
Plaque assays for titrating dengue virus (DENV) are time-consuming and not suitable for strains that do not plaque. Fluorescence-activated cell sorting (FACS) has been used to detect DENV-infected cells. Here we describe a FACS-based assay for titrating DENV. We determined that at 24 h postinfection, the number of infected cells detected by FACS represented the first round of infection and therefore could be used as a readout of the number of infectious particles in the inoculum. When the titers of different laboratory and clinical strains of DENV were compared using FACS, plaque, and endpoint dilution assays, for most strains the FACS titers were comparable to titers obtained by plaque or endpoint dilution assays. The FACS assay is an improvement over the plaque assay because the infection period is reduced from 5 to 7 days to 24 h and the assay can be used to titrate clinical isolates that frequently do not form clear plaques on cell monolayers. The novel FACS-based methods described here will facilitate laboratory studies of dengue.