Name
Development of a VP6 Binding ELISA and Luminex bead-based Assays for Rotavirus-Specific IgA and IgG
Presenter
Jonathan Mandolo, Malawi Liverpool Wellcome Trust
Co-Author(s)
Jonathan Mandolo1,2, Kondwani C. Jambo1,2,4*, Khuzwayo C. Jere1,3,4*
1Malawi Liverpool Wellcome Programme, Blantyre, Malawi.
2Department of Clinical Sciences, Liverpool School of Tropical medicine, L3 5QA, Liverpool, UK.
3Clinical Infection, Microbiology and Immunology, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool, UK.
4Medical Laboratories Department, Kamuzu University of Health Sciences, Blantyre, Malawi.
Abstract Category
Combatting and Exploiting dsRNA viruses
Abstract
Background
Continuous screening for rotavirus-specific antibodies is vital for immunity assessment and public health efforts. Traditionally, rabbit anti-rotavirus IgG (Capture ELISA) has been in use on rotavirus-specific antibodies screening. However, its discontinued production by key suppliers has a potential to disrupt the screening efforts globally. While effective, Capture ELISA requires specialized expertise and is labor-intensive. This study evaluates recombinant rotavirus A VP6 antigen on ELISA and Luminex bead-based platforms as scalable alternatives.
Methods
Recombinant rotavirus VP6 (CSB) was optimized and validated for detecting rotavirus-specific antibodies using ELISA and Luminex bead-based assays. Rabbit anti-rotavirus IgG-based Capture ELISA was the gold standard, with a positivity cutoff of rotavirus-specific IgA ≥ 20 IU/mL. Pearson’s correlation assessed the performance across platforms.
Results
Strong correlations were observed between IgA and IgG titers across platforms. The Capture ELISA and CSB ELISA showed nearly perfect correlation (IgA: R = 0.97, IgG: R = 0.95), as did the Capture ELISA and CSB Luminex platform (IgA: R = 0.98, IgG: R = 0.94), and the CSB ELISA and Luminex platform (IgA: R = 0.98, IgG: R = 0.94). The CSB ELISA showed 97.30% sensitivity, 100% specificity, and 98.41% concordance. The Luminex platform exhibited perfect sensitivity, specificity, and concordance.
Conclusion
Recombinant rotavirus VP6 on both ELISA and Luminex platforms provides reliable, scalable methods for detecting rotavirus-specific antibodies, offering efficient tools for large-scale immunity screening and global surveillance.
Continuous screening for rotavirus-specific antibodies is vital for immunity assessment and public health efforts. Traditionally, rabbit anti-rotavirus IgG (Capture ELISA) has been in use on rotavirus-specific antibodies screening. However, its discontinued production by key suppliers has a potential to disrupt the screening efforts globally. While effective, Capture ELISA requires specialized expertise and is labor-intensive. This study evaluates recombinant rotavirus A VP6 antigen on ELISA and Luminex bead-based platforms as scalable alternatives.
Methods
Recombinant rotavirus VP6 (CSB) was optimized and validated for detecting rotavirus-specific antibodies using ELISA and Luminex bead-based assays. Rabbit anti-rotavirus IgG-based Capture ELISA was the gold standard, with a positivity cutoff of rotavirus-specific IgA ≥ 20 IU/mL. Pearson’s correlation assessed the performance across platforms.
Results
Strong correlations were observed between IgA and IgG titers across platforms. The Capture ELISA and CSB ELISA showed nearly perfect correlation (IgA: R = 0.97, IgG: R = 0.95), as did the Capture ELISA and CSB Luminex platform (IgA: R = 0.98, IgG: R = 0.94), and the CSB ELISA and Luminex platform (IgA: R = 0.98, IgG: R = 0.94). The CSB ELISA showed 97.30% sensitivity, 100% specificity, and 98.41% concordance. The Luminex platform exhibited perfect sensitivity, specificity, and concordance.
Conclusion
Recombinant rotavirus VP6 on both ELISA and Luminex platforms provides reliable, scalable methods for detecting rotavirus-specific antibodies, offering efficient tools for large-scale immunity screening and global surveillance.