Zhang Yingze, Li Xiaoyun, Di Y Peter
Department of Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA, USA.
Methods Mol Biol. 2020;2102:129-147. doi: 10.1007/978-1-0716-0223-2_6.
Immunoassay is one of the most commonly used biomedical techniques to detect the expression of an antibody or an antigen in a test sample. Enzyme-linked immunosorbent assay (ELISA) has been used for a variety of applications including diagnostic tools and quality controls. However, one of the main limitations of ELISA is its lack of multiplexing ability, so ELISA may not be an efficient diagnostic tool when a measurement of multiple determinants is needed for samples with limited quantity such as blood or biological samples from newborns or babies. Although similar to ELISA in assay measurement, multiplex platforms such as bead-based Luminex and multi-array-based MSD (Meso Scale Discovery) are widely used to measure multiple biomarkers from a single analysis. Luminex is a xMAP-based technology that combines several different technologies to provide an efficient and accurate measurement of multiple analytes from a single sample. The multiplexing can be achieved because up to 100 distinct Luminex color-coded microsphere bead sets can be coated with a reagent specific to a particular bioassay, allowing the capture and detection of specific analytes from a sample. Using Multi-array and electrochemiluminescence technologies, the MSD platform provides the multiplex capability with similar consistence as observed in ELISA. Various biological samples that can be analyzed by both Luminex and MSD systems include serum, plasma, tissue and cell lysate, saliva, sputum, and bronchoalveolar Lavage (BAL). The most common Luminex and MSD-based assays are to detect a combined set of cytokines to provide a measurement of cytokine expression profiling for a diagnostic purpose.
免疫测定是检测测试样品中抗体或抗原表达的最常用生物医学技术之一。酶联免疫吸附测定（ELISA）已用于包括诊断工具和质量控制在内的各种应用。然而，ELISA的主要局限性之一是其缺乏多重检测能力，因此当需要对新生儿或婴儿的血液或生物样品等有限量样品进行多种决定因素的测量时，ELISA可能不是一种有效的诊断工具。尽管在测定测量方面与ELISA相似，但基于微珠的Luminex和基于多阵列的MSD（Meso Scale Discovery）等多重检测平台被广泛用于从单次分析中测量多种生物标志物。Luminex是一种基于xMAP的技术，它结合了几种不同的技术，能够从单个样品中高效、准确地测量多种分析物。之所以能够实现多重检测，是因为多达100种不同的Luminex颜色编码微球珠组可以包被特定生物测定的试剂，从而能够从样品中捕获和检测特定的分析物。MSD平台利用多阵列和电化学发光技术，提供了与ELISA相似的一致性的多重检测能力。Luminex和MSD系统都可以分析的各种生物样品包括血清、血浆、组织和细胞裂解物、唾液、痰液和支气管肺泡灌洗（BAL）液。最常见的基于Luminex和MSD的测定是检测一组组合的细胞因子，以提供用于诊断目的的细胞因子表达谱测量。
