Multiplexed particle-based flow cytometric assays
Multiplexed particle-based flow cytometric assays
复制标题
DOI：
10.1016/s0022-1759(00)00238-6
复制
发表时间：
2000-09-21
影响因子：
2.2
通讯作者：
Vignali, DAA
中科院分区：
文献类型：
--
作者：
Vignali, DAA
已经开发了几种方法来定量生物流体和组织培养样品中的可溶性分析物，包括生物测定、ELISA、RPA和PCR。然而，这些技术中的每一种都具有一个或多个显著的局限性; ELISA一次只能测量一种分析物; PCR不能检测天然蛋白质。基于颗粒的流式细胞术检测的最新发展带来了希望，许多这些限制可以克服。该技术利用微球作为常规免疫测定、亲和测定或DNA杂交测定的固体支持物，随后在流式细胞仪上进行分析。几种多重珠系统目前由不同的供应商销售。我们使用Luminex FlowMetrologTM系统，其由64种不同的珠组组成，所述珠组用均匀、不同比例的红色和橙子荧光染料（通过FACScan上的FL 2/FL 3检测）制造。每个微珠组构成了使用绿色荧光报告染料（FL 1）的单个检测的基础。该系统有助于开发多路检测，同时测量许多不同的分析物在一个小的样品体积。它们也可以发展成快速的、“无洗涤”的测定，
Several methods have been developed to quantify soluble analytes in biological fluids and tissue culture samples, including bioassays, ELISA, RPA and PCR. However, each of these techniques possesses one or more significant limitations; ELISA will only measure one analyte as a time; PCR does not detect native protein. The recent development of particle-based flow cytometric assays has raised hopes that many of these limitations can be overcome. The technology utilizes microspheres as the solid support for a conventional immunoassay, affinity assay or DNA hybridization assay which are subsequently analyzed on a flow cytometer. Several multiplexed bead systems are currently marketed by different vendors, We have used the Luminex FlowMetrix(TM) system which consists of 64 different bead sets manufactured with uniform, distinct proportions of red and orange fluorescent dyes (detected by FL2/FL3 on a FACScan). Each bead set forms the basis of an individual assay using a green fluorescent reporter dye (FL1). This system facilitates the development of multiplexed assays that simultaneously measure many different analytes in a small sample volume. They can also be developed into rapid, 'no wash' assays that can be completed in