优化多路复用高通量测定法，以测定针对 SARS-CoV-2 变体的中和抗体。

IF 2.2 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods Pub Date : 2024-11-16 DOI:10.1016/j.jviromet.2024.115073

Caroline L. Ashley , Malik Bloul , Sibel Alca , Lachlan Smith , Wang Jin , David Khoury , Claudio Counoupas , Miles Davenport , James A. Triccas , Megan Steain

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引用次数: 0

摘要

开发了一种基于慢病毒的多重伪病毒中和试验（pVNT），用于高通量测定针对三种不同的 SARS-CoV-2 尖峰变体的中和抗体（nAbs）。通过优化平板布局和确定伪病毒接种体的最佳转导百分比，最大限度地减少了测定内变异性。比较单一和多重 pVNT 检测的 EC50 滴度没有发现明显差异，这表明多重检测是可靠的。对康复期人类血清的评估证实了测定的稳健性，在单一和多重测定中观察到的变异伪病毒相对于祖先毒株的 EC50 滴度是一致的。这种多重 pVNT 为评估针对 SARS-CoV-2 变异株的 nAb 反应提供了可靠的工具，可用于加快临床前疫苗评估，为下一次冠状病毒大流行做准备。

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Optimisation of a multiplexed, high throughput assay to measure neutralising antibodies against SARS-CoV-2 variants

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 EC₅₀ 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 EC₅₀ 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.

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来源期刊

Journal of virological methods 医学-病毒学

CiteScore

5.80

自引率

0.00%

发文量

209

审稿时长

41 days

期刊介绍： The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.