da Rocha Queiroz Lima Monique, Monteiro-Maia Renata, Nunes Priscila Conrado Guerra
Strategic Laboratory for Molecular Diagnostics, Butantan Institute, São Paulo, Brazil.
Laboratory of Molecular Biology of Insects, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro, RJ, Brazil.
Methods Mol Biol. 2025;2913:239-246. doi: 10.1007/978-1-0716-4458-4_22.
The Mac-ELISA technique is based on the capture of IgM antibodies by the formation of an antigen-antibody complex in biological samples. IgM anti-yellow fever antibodies are captured by anti-IgM present on the polystyrene surface of the wells of the assay plate.The steps of the technique consist of stages (serum, antigen and anti-flavivirus conjugate - 6B6C-1), with incubation intervals at 37°C and washing. When adding the substrate in the next step, positive reactions (those containing the antigen-antibody complex) are blue in color by the action of the TMB substrate on the peroxidase enzyme. Completion of the reaction by adding the stop solution will cause the positive reaction of blue color to turn yellow.
Mac-ELISA技术基于在生物样品中通过形成抗原-抗体复合物来捕获IgM抗体。IgM抗黄热病毒抗体被存在于检测板孔聚苯乙烯表面的抗IgM捕获。该技术步骤包括多个阶段（血清、抗原和抗黄病毒缀合物-6B6C-1），在37℃下孵育并进行洗涤。在下一步添加底物时，阳性反应（那些含有抗原-抗体复合物的反应）由于TMB底物对过氧化物酶的作用而呈蓝色。加入终止液完成反应后，蓝色的阳性反应将变为黄色。
