A semi-automated ELISA workflow for SARS-CoV-2 Nucleocapsid IgG detection significantly improves throughput and reduces hands-on time

IF 1.7 4区医学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of immunological methods Pub Date : 2026-07-01 Epub Date: 2026-06-07 DOI:10.1016/j.jim.2026.114075

Ruturaj R. Masvekar, Gabriela Mansano Do Nascimento, Aliyah Hannah A. Remoroza, Shweta Jain, Troy J. Kemp, Bruce K. Brown, Ligia A. Pinto

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引用次数: 0

Abstract

Background

Enzyme-linked immunosorbent assay (ELISA) is widely used in clinical and research laboratories for applications such as disease surveillance, immunogenicity assessment, and vaccine evaluation. Traditional manual ELISA workflows, however, are labor-intensive, time-consuming, and susceptible to operator-dependent variability, which can limit throughput and reproducibility. Incorporating semi-automation into established protocols offers a potential strategy to improve efficiency while maintaining analytical performance.

Methods

A validated manual ELISA protocol for SARS-CoV-2 Nucleocapsid IgG detection was adapted into a semi-automated workflow using an automated multichannel pipette, microplate washer-dispenser, and plate stacker to streamline key steps. Performance was compared across 10 manual and 30 semi-automated plates. Sixteen pre-characterized serum samples were analyzed to evaluate precision, accuracy, and overall assay validity, and internal controls were monitored to ensure compliance with predefined acceptance criteria.

Results

The semi-automated workflow doubled assay throughput from 6 to 12 plates per analyst per day and reduced hands-on time by 75% (from 5.5 h to 1 h). Internal controls for all 40 plates met established acceptance ranges. Precision, measured as relative standard deviation, remained below 25% for all samples, and accuracy, expressed as percent error, remained below 20%. No loss of analytical performance was observed when transitioning from manual to semi-automated processing.

Conclusion

Semi-automation markedly increases ELISA throughput and reduces hands-on time while maintaining precision, accuracy, and overall assay validity. These results underscore its feasibility as an efficient workflow enhancement with potential for broader application across diverse ELISA platforms pending further validation.

查看原文本刊更多论文

用于SARS-CoV-2 IgG检测的半自动化ELISA工作流程显着提高了吞吐量并减少了操作时间。

背景：酶联免疫吸附测定（ELISA）广泛应用于临床和研究实验室，如疾病监测、免疫原性评估和疫苗评价。然而，传统的手工ELISA工作流程是劳动密集型的，耗时的，并且易受操作人员差异的影响，这可能会限制吞吐量和可重复性。将半自动化集成到已建立的协议中提供了一种潜在的策略，可以在保持分析性能的同时提高效率。方法：将经过验证的手工ELISA检测SARS-CoV-2核衣壳IgG的方案调整为半自动化的工作流程，使用自动化多通道移液器、微孔板洗涤分配器和叠板机，以简化关键步骤。对10个手动和30个半自动平板的性能进行了比较。对16份预表征血清样本进行分析，以评估精密度、准确性和总体测定效度，并监测内部控制以确保符合预定义的接受标准。结果：半自动化的工作流程使每位分析人员每天的分析通量从6个增加到12个，减少了75%的动手时间（从4 h减少到1 h）。所有40个板材的内部控制均符合既定验收范围。以相对标准偏差衡量的精密度在所有样本中保持在25%以下，以百分比误差表示的准确度保持在20%以下。从手动过渡到半自动处理时，没有观察到分析性能的损失。结论：半自动化显著提高了ELISA通量，减少了操作时间，同时保持了精密度、准确性和整体检测效度。这些结果强调了其作为一种有效的工作流程增强的可行性，并具有在不同ELISA平台上更广泛应用的潜力，有待进一步验证。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of immunological methods 医学-免疫学

CiteScore

4.10

自引率

0.00%

发文量

120

审稿时长

3 months

期刊介绍： The Journal of Immunological Methods is devoted to covering techniques for: (1) Quantitating and detecting antibodies and/or antigens. (2) Purifying immunoglobulins, lymphokines and other molecules of the immune system. (3) Isolating antigens and other substances important in immunological processes. (4) Labelling antigens and antibodies. (5) Localizing antigens and/or antibodies in tissues and cells. (6) Detecting, and fractionating immunocompetent cells. (7) Assaying for cellular immunity. (8) Documenting cell-cell interactions. (9) Initiating immunity and unresponsiveness. (10) Transplanting tissues. (11) Studying items closely related to immunity such as complement, reticuloendothelial system and others. (12) Molecular techniques for studying immune cells and their receptors. (13) Imaging of the immune system. (14) Methods for production or their fragments in eukaryotic and prokaryotic cells. In addition the journal will publish articles on novel methods for analysing the organization, structure and expression of genes for immunologically important molecules such as immunoglobulins, T cell receptors and accessory molecules involved in antigen recognition, processing and presentation. Submitted full length manuscripts should describe new methods of broad applicability to immunology and not simply the application of an established method to a particular substance - although papers describing such applications may be considered for publication as a short Technical Note. Review articles will also be published by the Journal of Immunological Methods. In general these manuscripts are by solicitation however anyone interested in submitting a review can contact the Reviews Editor and provide an outline of the proposed review.