优化一种用于检测针对新冠病毒变异株的中和抗体的多重高通量检测方法。
Optimisation of a multiplexed, high throughput assay to measure neutralising antibodies against SARS-CoV-2 variants.
作者信息
Ashley Caroline L, Bloul Malik, Alca Sibel, Smith Lachlan, Jin Wang, Khoury David, Counoupas Claudio, Davenport Miles, Triccas James A, Steain Megan
机构信息
Sydney Infectious Diseases Institute (Sydney ID), Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; Charles Perkin Centre, The University of Sydney, Camperdown, NSW 2006, Australia.
Sydney Infectious Diseases Institute (Sydney ID), Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; Charles Perkin Centre, The University of Sydney, Camperdown, NSW 2006, Australia.
出版信息
J Virol Methods. 2025 Feb;332:115073. doi: 10.1016/j.jviromet.2024.115073. Epub 2024 Nov 16.
A multiplexed, lentivirus-based pseudovirus neutralisation assay (pVNT) was developed for high-throughput measurement of neutralising antibodies (nAbs) against three distinct SARS-CoV-2 spike variants. Intra-assay variability was minimised by optimising the plate layout and determining an optimal percentage transduction for the pseudovirus inoculum. Comparison of EC titres between single and multiplexed pVNT assays showed no significant differences, indicating reliability of the multiplexed assay. Evaluation of convalescent human sera confirmed assay robustness, with consistent EC titres for variant pseudoviruses relative to the ancestral strain observed across single and multiplexed assays. This multiplexed pVNT provides a reliable tool for assessing nAb responses against SARS-CoV-2 variants and could be used to accelerate preclinical vaccine assessment in preparation for the next coronavirus pandemic.
开发了一种基于慢病毒的多重假病毒中和试验（pVNT），用于高通量测量针对三种不同的SARS-CoV-2刺突变体的中和抗体（nAbs）。通过优化板布局和确定假病毒接种物的最佳转导百分比，将试验内变异性降至最低。单重和多重pVNT试验之间的EC滴度比较显示无显著差异，表明多重试验的可靠性。对康复期人类血清的评估证实了试验的稳健性，在单重和多重试验中观察到，相对于原始毒株，变异假病毒的EC滴度一致。这种多重pVNT为评估针对SARS-CoV-2变体的nAb反应提供了一种可靠的工具，可用于加速临床前疫苗评估，为下一次冠状病毒大流行做好准备。
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