Ahsan Haseeb
Department of Biochemistry, Faculty of Dentistry, Jamia Millia Islamia (A Central University), New Delhi - 110025, India.
Comp Clin Path. 2022;31(2):333-345. doi: 10.1007/s00580-021-03302-4. Epub 2021 Nov 20.
Immunoassays are a powerful diagnostic tool and are widely used for the quantification of proteins and biomolecules in medical diagnosis and research. Enzyme-linked immunosorbent assay (ELISA) is the most commonly used immunoassay format and allows the detection of biomarkers at a very low concentration. The diagnostic platforms such as enzyme immunoassay (EIA), chemiluminescence (CL) assay, polymerase chain reaction (PCR), flow cytometry (FC), and mass spectrometry (MS) have been used to identify molecular biomarkers. However, these diagnostic tools requiring expensive equipment, long testing time, and qualified personnel that is not always available in small local hospitals with limited resources. The lateral flow immunoassay (LFIA) platform was developed for rapidly obtaining laboratory results and to make urgent decisions in emergency medicine, as well as the recently introduced concept of testing at the site of care (point-of-care, POC). The simultaneous measurement of different substances from a single sample called multiplex assays have become increasingly significant for in vitro quantification of multiple analytes in a single sample, thereby minimising cost, time, and volume. In multiplex immunoassays, the ligands are immobilized either in planar format (flat surface) or on microspheres in suspension that binds to target analytes in sample. The multiplex technology has established itself in proteomic networks and pathways, validation of genomic discoveries, and in the development of clinical biomarkers. In the present review article, various types of monoplex/simplex and complex/multiplex immunoassays have been analysed that are increasingly being applied in laboratory medicine. Also, some advantages and disadvantages of these multiplex assays have also been included such as experimental animals, in vitro tests using cell lines and tissue samples, 3-dimensional modelling and bioprinting, in silico tests, organ-on-chip, and computer modelling.
免疫测定是一种强大的诊断工具，广泛用于医学诊断和研究中蛋白质和生物分子的定量分析。酶联免疫吸附测定（ELISA）是最常用的免疫测定形式，能够检测极低浓度的生物标志物。酶免疫测定（EIA）、化学发光（CL）测定、聚合酶链反应（PCR）、流式细胞术（FC）和质谱（MS）等诊断平台已被用于识别分子生物标志物。然而，这些诊断工具需要昂贵的设备、较长的检测时间，并且需要合格的人员，而资源有限的小型地方医院不一定总能满足这些要求。横向流动免疫测定（LFIA）平台的开发是为了在急诊医学中快速获得实验室结果并做出紧急决策，以及最近引入的现场检测（即时检测，POC）概念。从单个样品中同时测量不同物质的多重分析对于在单个样品中体外定量多种分析物变得越来越重要，从而最大限度地降低成本、时间和体积。在多重免疫测定中，配体以平面形式（平面表面）或悬浮在微球上的形式固定，与样品中的目标分析物结合。多重技术已在蛋白质组网络和途径、基因组发现的验证以及临床生物标志物的开发中确立了自身地位。在本综述文章中，分析了越来越多地应用于检验医学的各种类型的单重/单纯和复杂/多重免疫测定。此外，还包括了这些多重分析的一些优缺点，如实验动物、使用细胞系和组织样品的体外试验、三维建模和生物打印、计算机模拟试验、芯片器官和计算机建模。
