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  • MultiSero: An Open-Source Multiplex-ELISA Platform for Measuring Antibody Responses to Infection. Pathogens (Basel, Switzerland) Byrum, J. R., Waltari, E., Janson, O., Guo, S., Folkesson, J., Chhun, B. B., Vinden, J., Ivanov, I. E., Forst, M. L., Li, H., Larson, A. G., Blackmon, L., Liu, Z., Wu, W., Ahyong, V., Tato, C. M., McCutcheon, K. M., Hoh, R., Kelly, J. D., Martin, J. N., Peluso, M. J., Henrich, T. J., Deeks, S. G., Prakash, M., Greenhouse, B., Mehta, S. B., Pak, J. E. 2023; 12 (5)


    A multiplexed enzyme-linked immunosorbent assay (ELISA) that simultaneously measures antibody binding to multiple antigens can extend the impact of serosurveillance studies, particularly if the assay approaches the simplicity, robustness, and accuracy of a conventional single-antigen ELISA. Here, we report on the development of multiSero, an open-source multiplex ELISA platform for measuring antibody responses to viral infection. Our assay consists of three parts: (1) an ELISA against an array of proteins in a 96-well format; (2) automated imaging of each well of the ELISA array using an open-source plate reader; and (3) automated measurement of optical densities for each protein within the array using an open-source analysis pipeline. We validated the platform by comparing antibody binding to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) antigens in 217 human sera samples, showing high sensitivity (0.978), specificity (0.977), positive predictive value (0.978), and negative predictive value (0.977) for classifying seropositivity, a high correlation of multiSero determined antibody titers with commercially available SARS-CoV-2 antibody tests, and antigen-specific changes in antibody titer dynamics upon vaccination. The open-source format and accessibility of our multiSero platform can contribute to the adoption of multiplexed ELISA arrays for serosurveillance studies, for SARS-CoV-2 and other pathogens of significance.

    View details for DOI 10.3390/pathogens12050671

    View details for PubMedID 37242341