Development of a novel multiplex electrochemiluminescent-based immunoassay for quantification of human serum IgG against 10 Staphylococcus aureus toxins

详细信息    查看全文

• 作者：Rajan P. Adhikari ; ^{rajan@integratedbiotherapeutics.com" class="auth_mail" title="E-mail the corresponding author} ; Christian Haudenschild ; Patricia M. Sterba ; Sara Sahandi ; Sven Enterlein ; Frederick W. Holtsberg ; M. Javad Aman
• 关键词：Multiplex ; ELISA serology ; Staphylococcal exotoxins ; Delta toxin
• 刊名：Journal of Immunological Methods
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：430
• 期：Complete
• 页码：33-42
• 全文大小：1555 K

文摘

An electrochemiluminescent (ECL)-based multiplex immunoassay using Meso-Scale Discovery (MSD) technology was developed for detecting antibody response toward 10 Staphylococcus aureus (S. aureus) exotoxins. These 10 antigens included three different groups of toxins: 1) single component pore-forming toxins such as alpha- and delta-hemolysins, 2) the bicomponent pore-forming toxin Panton–Valentine leukocidin (PVL), comprised of LukS-PV and LukF-PV subunits, and 3) enterotoxin/superantigens — Staphylococcal enterotoxins A (SEA), B (SEB), C1 (SEC1), D (SED), K (SEK) and Toxic shock syndrome toxin-1 (TSST-1). Assay development included optimization steps with a conventional SEB ELISA-based serological assay and then optimized parameters were transferred and re-optimized in a singleplex ECL format. Finally, two pentaplex solid-phase ECL formats were developed. As proof of concept, one set of pentaplex ECL data was compared with conventional ELISA results. During the assay development controls were screened and developed for both the singleplex and multiplex assays. ECL-based multiplex assays were more sensitive with a wide dynamic range and proved more time-efficient than conventional ELISAs. Using the newly developed ECL method we showed, for the first time, that delta-hemolysin toxin can induce an immune response as antibody titers could be detected.