Gerald Waweru, Ruth Nyakundi, Bernadette Kutima, Sharon Owuor, Gloria Konyino, John Gitonga, Doreen Lugano, Angela Maina, Jennifer Musyoki, Lucy Ochola, Martin Omondi, Christopher K Kariuki, Paul Ogongo, Christina Mwachari, Faiz Shee, Charles Agoti, Charles Sande, Sophie Uyoga, Eunice Kagucia, Ambrose Agweyu, Philip Bejon, J Anthony G Scott, George M Warimwe, L Isabella Ochola-Oyier, James Nyagwange
{"title":"评估SARS-CoV-2刺突突变对抗体结合的影响:在多重Luminex试验中对武汉和JN.1变异全长刺突的比较评估","authors":"Gerald Waweru, Ruth Nyakundi, Bernadette Kutima, Sharon Owuor, Gloria Konyino, John Gitonga, Doreen Lugano, Angela Maina, Jennifer Musyoki, Lucy Ochola, Martin Omondi, Christopher K Kariuki, Paul Ogongo, Christina Mwachari, Faiz Shee, Charles Agoti, Charles Sande, Sophie Uyoga, Eunice Kagucia, Ambrose Agweyu, Philip Bejon, J Anthony G Scott, George M Warimwe, L Isabella Ochola-Oyier, James Nyagwange","doi":"10.3390/v17091248","DOIUrl":null,"url":null,"abstract":"<p><p>Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) continues to evolve, with mutations leading to the emergence of new variants. JN.1, a subvariant of omicron BA.2.86, has demonstrated marked immune escape and is now included in updated vaccine formulations. While reduced sensitivity has been reported for antibody assays using ancestral spike protein subunits to detect omicron-induced responses, the performance of full-length spike-based assays against omicron sublineages remains unclear. We aimed to compare the sensitivity of ELISA and Luminex assays using full-length spike proteins from the ancestral Wuhan strain and the JN.1 variant.</p><p><strong>Methods: </strong>Wuhan and JN.1 full-length spike protein constructs were designed and expressed in Expi293F mammalian cells. In-house ELISAs based on previously validated protocols were used to measure anti-spike IgG levels. Additionally, a Luminex-based assay for anti-spike antibody detection was developed and validated. Both assays were applied to the following sample groups: pre-pandemic samples (designated \"gold standard negatives\"); PCR confirmed 2020 positives (\"gold standard wildtype positives\"); PCR confirmed 2024 positives (\"gold standard omicron positives\"); 2022 vaccinated individuals with verbal confirmed infection (\"gold standard hybrid positives\"); and 2024 household samples (\"unknowns\").</p><p><strong>Results: </strong>Wuhan spike protein showed a sensitivity of 100% (95% CI: 0.88-1.0) in detecting omicron-specific antibodies using gold standard omicron positives with JN.1 spike protein as a reference assay. Overall, across all samples, in ELISA, the Wuhan antigen had a sensitivity of 0.93 (95% CI: 0.89-0.95) and a specificity of 0.98 (95% CI: 0.94-0.99). The JN.1 antigen showed a sensitivity of 0.91 (95% CI: 0.87-0.94) and a specificity of 0.97 (95% CI: 0.93-0.99). In Luminex, sensitivity was 0.95 (95% CI: 0.91-0.97) for Wuhan and 0.94 (95% CI: 0.91-0.96) for JN.1. Specificity for both antigens in Luminex was 0.98 (95% CI: 0.94-0.99).</p><p><strong>Conclusions: </strong>Both ELISA and Luminex assays showed comparable sensitivity and specificity for both Wuhan and JN.1 antigens, indicating that mutations in the JN.1 variant do not significantly impact assay performance. This suggests preserved antigenic recognition across variants.</p>","PeriodicalId":49328,"journal":{"name":"Viruses-Basel","volume":"17 9","pages":""},"PeriodicalIF":3.5000,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12474121/pdf/","citationCount":"0","resultStr":"{\"title\":\"Assessing the Impact of SARS-CoV-2 Spike Mutations on Antibody Binding: A Comparative Assessment of the Wuhan and JN.1 Variants' Full-Length Spikes in a Multiplex Luminex Assay.\",\"authors\":\"Gerald Waweru, Ruth Nyakundi, Bernadette Kutima, Sharon Owuor, Gloria Konyino, John Gitonga, Doreen Lugano, Angela Maina, Jennifer Musyoki, Lucy Ochola, Martin Omondi, Christopher K Kariuki, Paul Ogongo, Christina Mwachari, Faiz Shee, Charles Agoti, Charles Sande, Sophie Uyoga, Eunice Kagucia, Ambrose Agweyu, Philip Bejon, J Anthony G Scott, George M Warimwe, L Isabella Ochola-Oyier, James Nyagwange\",\"doi\":\"10.3390/v17091248\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) continues to evolve, with mutations leading to the emergence of new variants. JN.1, a subvariant of omicron BA.2.86, has demonstrated marked immune escape and is now included in updated vaccine formulations. While reduced sensitivity has been reported for antibody assays using ancestral spike protein subunits to detect omicron-induced responses, the performance of full-length spike-based assays against omicron sublineages remains unclear. We aimed to compare the sensitivity of ELISA and Luminex assays using full-length spike proteins from the ancestral Wuhan strain and the JN.1 variant.</p><p><strong>Methods: </strong>Wuhan and JN.1 full-length spike protein constructs were designed and expressed in Expi293F mammalian cells. In-house ELISAs based on previously validated protocols were used to measure anti-spike IgG levels. Additionally, a Luminex-based assay for anti-spike antibody detection was developed and validated. Both assays were applied to the following sample groups: pre-pandemic samples (designated \\\"gold standard negatives\\\"); PCR confirmed 2020 positives (\\\"gold standard wildtype positives\\\"); PCR confirmed 2024 positives (\\\"gold standard omicron positives\\\"); 2022 vaccinated individuals with verbal confirmed infection (\\\"gold standard hybrid positives\\\"); and 2024 household samples (\\\"unknowns\\\").</p><p><strong>Results: </strong>Wuhan spike protein showed a sensitivity of 100% (95% CI: 0.88-1.0) in detecting omicron-specific antibodies using gold standard omicron positives with JN.1 spike protein as a reference assay. Overall, across all samples, in ELISA, the Wuhan antigen had a sensitivity of 0.93 (95% CI: 0.89-0.95) and a specificity of 0.98 (95% CI: 0.94-0.99). The JN.1 antigen showed a sensitivity of 0.91 (95% CI: 0.87-0.94) and a specificity of 0.97 (95% CI: 0.93-0.99). In Luminex, sensitivity was 0.95 (95% CI: 0.91-0.97) for Wuhan and 0.94 (95% CI: 0.91-0.96) for JN.1. Specificity for both antigens in Luminex was 0.98 (95% CI: 0.94-0.99).</p><p><strong>Conclusions: </strong>Both ELISA and Luminex assays showed comparable sensitivity and specificity for both Wuhan and JN.1 antigens, indicating that mutations in the JN.1 variant do not significantly impact assay performance. This suggests preserved antigenic recognition across variants.</p>\",\"PeriodicalId\":49328,\"journal\":{\"name\":\"Viruses-Basel\",\"volume\":\"17 9\",\"pages\":\"\"},\"PeriodicalIF\":3.5000,\"publicationDate\":\"2025-09-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12474121/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Viruses-Basel\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.3390/v17091248\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"VIROLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Viruses-Basel","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3390/v17091248","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"VIROLOGY","Score":null,"Total":0}
Assessing the Impact of SARS-CoV-2 Spike Mutations on Antibody Binding: A Comparative Assessment of the Wuhan and JN.1 Variants' Full-Length Spikes in a Multiplex Luminex Assay.
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) continues to evolve, with mutations leading to the emergence of new variants. JN.1, a subvariant of omicron BA.2.86, has demonstrated marked immune escape and is now included in updated vaccine formulations. While reduced sensitivity has been reported for antibody assays using ancestral spike protein subunits to detect omicron-induced responses, the performance of full-length spike-based assays against omicron sublineages remains unclear. We aimed to compare the sensitivity of ELISA and Luminex assays using full-length spike proteins from the ancestral Wuhan strain and the JN.1 variant.
Methods: Wuhan and JN.1 full-length spike protein constructs were designed and expressed in Expi293F mammalian cells. In-house ELISAs based on previously validated protocols were used to measure anti-spike IgG levels. Additionally, a Luminex-based assay for anti-spike antibody detection was developed and validated. Both assays were applied to the following sample groups: pre-pandemic samples (designated "gold standard negatives"); PCR confirmed 2020 positives ("gold standard wildtype positives"); PCR confirmed 2024 positives ("gold standard omicron positives"); 2022 vaccinated individuals with verbal confirmed infection ("gold standard hybrid positives"); and 2024 household samples ("unknowns").
Results: Wuhan spike protein showed a sensitivity of 100% (95% CI: 0.88-1.0) in detecting omicron-specific antibodies using gold standard omicron positives with JN.1 spike protein as a reference assay. Overall, across all samples, in ELISA, the Wuhan antigen had a sensitivity of 0.93 (95% CI: 0.89-0.95) and a specificity of 0.98 (95% CI: 0.94-0.99). The JN.1 antigen showed a sensitivity of 0.91 (95% CI: 0.87-0.94) and a specificity of 0.97 (95% CI: 0.93-0.99). In Luminex, sensitivity was 0.95 (95% CI: 0.91-0.97) for Wuhan and 0.94 (95% CI: 0.91-0.96) for JN.1. Specificity for both antigens in Luminex was 0.98 (95% CI: 0.94-0.99).
Conclusions: Both ELISA and Luminex assays showed comparable sensitivity and specificity for both Wuhan and JN.1 antigens, indicating that mutations in the JN.1 variant do not significantly impact assay performance. This suggests preserved antigenic recognition across variants.
期刊介绍:
Viruses (ISSN 1999-4915) is an open access journal which provides an advanced forum for studies of viruses. It publishes reviews, regular research papers, communications, conference reports and short notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. We also encourage the publication of timely reviews and commentaries on topics of interest to the virology community and feature highlights from the virology literature in the ''News and Views'' section. Electronic files or software regarding the full details of the calculation and experimental procedure, if unable to be published in a normal way, can be deposited as supplementary material.