Evaluation and deployment of isotype-specific salivary antibody assays for detecting previous SARS-CoV-2 infection in children and adults.
Evaluation and deployment of isotype-specific salivary antibody assays for detecting previous SARS-CoV-2 infection in children and adults.
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DOI：
10.1038/s43856-023-00264-2
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发表时间：
2023-03-15
期刊：
影响因子：
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通讯作者：
Halliday, Alice
中科院分区：
文献类型：
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作者：
Thomas, Amy C;Oliver, Elizabeth;Baum, Holly E;Gupta, Kapil;Shelley, Kathryn L;Long, Anna E;Jones, Hayley E;Smith, Joyce;Hitchings, Benjamin;di Bartolo, Natalie;Vasileiou, Kate;Rabi, Fruzsina;Alamir, Hanin;Eghleilib, Malak;Francis, Ore;Oliver, Jennifer;Morales-Aza, Begonia;Obst, Ulrike;Shattock, Debbie;Barr, Rachael;Collingwood, Lucy;Duale, Kaltun;Grace, Niall;Livera, Guillaume Gonnage;Bishop, Lindsay;Downing, Harriet;Rodrigues, Fernanda;Timpson, Nicholas;Relton, Caroline L;Toye, Ashley;Woolfson, Derek N;Berger, Imre;Goenka, Anu;Davidson, Andrew D;Gillespie, Kathleen M;Williams, Alistair J K;Bailey, Mick;Brooks-Pollock, Ellen;Finn, Adam;Halliday, Alice
唾液很容易非侵入性地获得，并且可能适用于检测当前和以前的SARS-CoV-2感染，但唾液抗体检测用于社区监测的证据有限。建立了唾液中SARS-CoV-2刺突蛋白、受体结合区域和核衣壳蛋白的IgA和IgG抗体检测试剂盒。我们评估了诊断性能，并使用配对的唾液和血清样本，相关的粘膜和全身抗体反应。在20个家庭暴发中对效果最好的检测方法进行了现场测试。我们证明了测试准确性（N = 320）， spike IgG （ROC AUC: 95.0%, 92.8-97.3%）和spike IgA （ROC AUC: 89.9%, 86.5-93.2%）检测可以最好地区分COVID-19大流行前和后的唾液样本。在年龄较小的年龄组（0-19岁）中，IgA和IgG的特异性为100%。然而，最佳检测方法的灵敏度较低（峰值IgG: 50.6%, 39.8-61.4%）。使用机器学习，当使用组合测试时，诊断性能得到改善。正如预期的那样，唾液IgA与血清相关性较差，表明口腔黏膜反应，而唾液IgG反应可预测血清反应。当部署到家庭暴发时，抗体反应是异质的，但仍然是最近感染的可靠指标。有趣的是，未接种疫苗但未确诊感染的儿童几乎完全通过特定的IgA反应暴露。通过强有力的标准化、评估和现场测试，这项工作为进一步研究SARS-CoV-2的传播和粘膜免疫提供了一个平台，并有可能将唾液监测扩大到难以到达的环境中的其他呼吸道感染。如果一个人以前感染过SARS-CoV-2，他们会产生特定的蛋白质，称为抗体。它们存在于唾液和血液中。唾液比血液更容易获得，因此我们开发并评估了六种检测儿童和成人唾液中SARS-CoV-2抗体的检测方法。一些测试检测到SARS-CoV-2产生的一种特定蛋白质的抗体，这种蛋白质被称为刺突蛋白，这些测试效果最好。最准确的结果是通过使用组合测试获得的。还可以开发类似的检测方法来检测其他呼吸道感染，从而更容易识别受感染的个体。Thomas等人用6种elisa检测唾液中SARS-CoV-2刺突蛋白、其受体结合区域和核衣壳蛋白的IgA和IgG抗体。在20起家庭疫情中，抗体反应各不相同，但却是近期感染的可靠指标。
Saliva is easily obtainable non-invasively and potentially suitable for detecting both current and previous SARS-CoV-2 infection, but there is limited evidence on the utility of salivary antibody testing for community surveillance. We established 6 ELISAs detecting IgA and IgG antibodies to whole SARS-CoV-2 spike protein, to its receptor binding domain region and to nucleocapsid protein in saliva. We evaluated diagnostic performance, and using paired saliva and serum samples, correlated mucosal and systemic antibody responses. The best-performing assays were field-tested in 20 household outbreaks. We demonstrate in test accuracy (N = 320), spike IgG (ROC AUC: 95.0%, 92.8–97.3%) and spike IgA (ROC AUC: 89.9%, 86.5–93.2%) assays to discriminate best between pre-pandemic and post COVID-19 saliva samples. Specificity was 100% in younger age groups (0–19 years) for spike IgA and IgG. However, sensitivity was low for the best-performing assay (spike IgG: 50.6%, 39.8–61.4%). Using machine learning, diagnostic performance was improved when a combination of tests was used. As expected, salivary IgA was poorly correlated with serum, indicating an oral mucosal response whereas salivary IgG responses were predictive of those in serum. When deployed to household outbreaks, antibody responses were heterogeneous but remained a reliable indicator of recent infection. Intriguingly, unvaccinated children without confirmed infection showed evidence of exposure almost exclusively through specific IgA responses. Through robust standardisation, evaluation and field-testing, this work provides a platform for further studies investigating SARS-CoV-2 transmission and mucosal immunity with the potential for expanding salivo-surveillance to other respiratory infections in hard-to-reach settings. If a person has been previously infected with SARS-CoV-2 they will produce specific proteins, called antibodies. These are present in the saliva and blood. Saliva is easier to obtain than blood, so we developed and evaluated six tests that detect SARS-CoV-2 antibodies in saliva in children and adults. Some tests detected antibodies to a particular protein made by SARS-CoV-2 called the spike protein, and these tests worked best. The most accurate results were obtained by using a combination of tests. Similar tests could also be developed to detect other respiratory infections which will enable easier identification of infected individuals. Thomas et al. test 6 ELISAs detecting IgA and IgG antibodies to whole SARS-CoV-2 spike protein, to its receptor binding domain region and to nucleocapsid protein in saliva. Across 20 household outbreaks, antibody responses are heterogeneous, but a reliable indicator of recent infection.