Abstract
Autoantibodies to cellular constituents are the serological hallmark in systemic autoimmune rheumatic diseases (1). They are often associated with certain diagnosis, clinical features, or disease activity and considered as clinically useful biomarkers. Autoantibody immunoassays have been used extensively for over 50 years with continuous changes in technologies and antigens used. While standard enzyme-linked immunosorbent assay (ELISA) is still commonly used in practice, migration to multiplex immunoassays is the direction that many companies and laboratories are moving toward. Although multiplex assays have many advantages over conventional assays, there are also problems that may cause confusions among clinicians and researchers. In this Opinion Article, advantages and current problems in the use of multiplex immunoassays are discussed.
Highlights
Line immunoassay (LIA) is somewhat similar to dot blot or western blot as a diluted serum is incubated with a strip that has several specific antigens in different areas on a strip
Beads with different sizes and/or fluorochromes with different colors or intensities are coated with different specific antigens and mixed to allow detection of each specific autoantibody by gating on beads with certain characteristic
The reasons that multiplex assays are replacing conventional enzyme-linked immunosorbent assay (ELISA) are to save time, material and labor cost, and allow efficient handling of a large number of samples to enhance the overall throughput for companies and laboratories
Summary
Multiplex autoantibody assays are ones that can detect many specific autoantibodies in a single run whereas a traditional ELISA uses a single antigen to detect only a single specificity of autoantibodies. Immunoassays using crude antigens are not generally considered as multiplex assays even though classical double immunodiffusion or immunoprecipitation can detect many specific autoantibodies in a single run. LIA is somewhat similar to dot blot or western blot as a diluted serum is incubated with a strip that has several specific antigens in different areas on a strip. Many different specific antigens are coated on a slide or a membrane These strips, mixture of beads, or a slide/membrane with multiple antigens is incubated with a diluted serum, and many specific autoantibodies can be determined simultaneously. The reasons that multiplex assays are replacing conventional ELISA are to save time, material and labor cost, and allow efficient handling of a large number of samples to enhance the overall throughput for companies and laboratories. While new multiplex immunoassays have certain advantages over conventional assays, using them with incomplete understanding, or without validation of the new assay against classic or standard assays, has caused many concerns, confusions, and problems in autoantibody immunoassays for clinicians and patients
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