Biosystems and Biomaterials Division, National Institute of Standards and Technology (NIST), Gaithersburg, MD 20899, USA.
Int J Mol Sci. 2023 Aug 2;24(15):12332. doi: 10.3390/ijms241512332.
COVID-19 is an ongoing, global pandemic caused by the novel, highly infectious SARS-CoV-2 virus. Efforts to mitigate the effects of SARS-CoV-2, such as mass vaccination and development of monoclonal therapeutics, require precise measurements of correlative, functional neutralizing antibodies that block virus infection. The development of rapid, safe, and easy-to-use neutralization assays is essential for faster diagnosis and treatment. Here, we developed a vesicular stomatitis virus (VSV)-based neutralization assay with two readout methods, imaging and flow cytometry, that were capable of quantifying varying degrees of neutralization in patient serum samples. We tested two different spike-pseudoviruses and conducted a time-course assay at multiple multiplicities of infection (MOIs) to optimize the assay workflow. The results of this assay correlate with the results of previously developed serology and surrogate neutralization assays. The two pseudovirus readout methods produced similar values of 50% neutralization titer values. Harvest-free in situ readouts for live-cell imaging and high-throughput analysis results for flow cytometry can provide unique capabilities for fast evaluation of neutralization, which is critical for the mitigation of future pandemics.
COVID-19 是由新型高传染性 SARS-CoV-2 病毒引起的持续全球大流行。减轻 SARS-CoV-2 影响的努力，如大规模疫苗接种和单克隆治疗药物的开发，需要精确测量相关的、具有功能的中和抗体来阻断病毒感染。开发快速、安全且易于使用的中和测定法对于更快的诊断和治疗至关重要。在这里，我们开发了一种基于水疱性口炎病毒（VSV）的中和测定法，具有两种读数方法，即成像和流式细胞术，能够定量测定患者血清样本中不同程度的中和作用。我们测试了两种不同的刺突假病毒，并在多个感染复数（MOI）下进行了时间过程测定，以优化测定工作流程。该测定法的结果与先前开发的血清学和替代中和测定法的结果相关。两种假病毒读数方法产生了相似的 50%中和效价值。用于活细胞成像的无收获原位读数和用于流式细胞术的高通量分析结果可以为快速评估中和作用提供独特的能力，这对于减轻未来的大流行至关重要。
