Moncunill Gemma, Campo Joseph J, Dobaño Carlota
Barcelona Centre for International Health Research (CRESIB), Hospital Clínic - Universitat de Barcelona, Carrer Roselló 153 (CEK building), E-08036, Barcelona, Catalonia, Spain,
Methods Mol Biol. 2014;1172:65-86. doi: 10.1007/978-1-4939-0928-5_6.
Quantitative suspension array technology allows the simultaneous measurement of different cytokines and chemokines in small sample volumes. The possibility of measuring multiple variables is important for discovery of biomarkers of pathogenesis or protection in complex diseases as well as measurement of antigen-specific cellular responses. Measurements can be made in biological specimens, such as plasma or serum, cell culture supernatants, and others. This technology is based on a capture-detection sandwich-type assay using fluorescent microspheres analyzable by Luminex instruments or flow cytometers. The complexity and cost of producing highly multiplexed cytokine/chemokine in-house assays make them especially apt for commercial production. There are several commercial kits available that vary in absolute cytokine concentration, sensitivity, reproducibility, and cost. This chapter gives an overview of cytometric bead array technology, introduces some of the kits, and provides detailed information about the one that performed well in a comparative study (Cytokine Human Magnetic 30-Plex Panel from Life Technologies™).
定量悬浮阵列技术能够在小样本体积中同时测量不同的细胞因子和趋化因子。对于发现复杂疾病发病机制或保护性生物标志物以及测量抗原特异性细胞反应而言，测量多个变量的可能性至关重要。测量可在生物标本中进行，如血浆、血清、细胞培养上清液等。该技术基于一种捕获 - 检测夹心型检测方法，使用可通过Luminex仪器或流式细胞仪分析的荧光微球。自行生产高度多重化的细胞因子/趋化因子检测方法的复杂性和成本使得它们特别适合商业化生产。有几种商业试剂盒可供选择，它们在绝对细胞因子浓度、灵敏度、重现性和成本方面各不相同。本章概述了细胞计数微珠阵列技术，介绍了一些试剂盒，并提供了在一项比较研究中表现良好的试剂盒（来自Life Technologies™的细胞因子人磁性30重检测板）的详细信息。
