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Development of a high-sensitivity ELISA detecting IgG, IgA and IgM antibodies to the SARS-CoV-2 spike glycoprotein in serum and saliva
Immunology ( IF 4.9 ) Pub Date : 2021-05-01 , DOI: 10.1111/imm.13349 Sian E Faustini 1 , Sian E Jossi 1 , Marisol Perez-Toledo 1 , Adrian M Shields 1 , Joel D Allen 2 , Yasunori Watanabe 2, 3 , Maddy L Newby 2 , Alex Cook 4 , Carrie R Willcox 1 , Mahboob Salim 1 , Margaret Goodall 1 , Jennifer L Heaney 1 , Edith Marcial-Juarez 1 , Gabriella L Morley 5 , Barbara Torlinska 6 , David C Wraith 1 , Tonny V Veenith 7 , Stephen Harding 4 , Stephen Jolles 8 , Mark J Ponsford 8 , Tim Plant 1 , Aarnoud Huissoon 1, 9 , Matthew K O'Shea 5 , Benjamin E Willcox 1 , Mark T Drayson 1 , Max Crispin 2 , Adam F Cunningham 1 , Alex G Richter 1
Immunology ( IF 4.9 ) Pub Date : 2021-05-01 , DOI: 10.1111/imm.13349 Sian E Faustini 1 , Sian E Jossi 1 , Marisol Perez-Toledo 1 , Adrian M Shields 1 , Joel D Allen 2 , Yasunori Watanabe 2, 3 , Maddy L Newby 2 , Alex Cook 4 , Carrie R Willcox 1 , Mahboob Salim 1 , Margaret Goodall 1 , Jennifer L Heaney 1 , Edith Marcial-Juarez 1 , Gabriella L Morley 5 , Barbara Torlinska 6 , David C Wraith 1 , Tonny V Veenith 7 , Stephen Harding 4 , Stephen Jolles 8 , Mark J Ponsford 8 , Tim Plant 1 , Aarnoud Huissoon 1, 9 , Matthew K O'Shea 5 , Benjamin E Willcox 1 , Mark T Drayson 1 , Max Crispin 2 , Adam F Cunningham 1 , Alex G Richter 1
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Detecting antibody responses during and after SARS-CoV-2 infection is essential in determining the seroepidemiology of the virus and the potential role of antibody in disease. Scalable, sensitive and specific serological assays are essential to this process. The detection of antibody in hospitalized patients with severe disease has proven relatively straightforward; detecting responses in subjects with mild disease and asymptomatic infections has proven less reliable. We hypothesized that the suboptimal sensitivity of antibody assays and the compartmentalization of the antibody response may contribute to this effect. We systematically developed an ELISA, optimizing different antigens and amplification steps, in serum and saliva from non-hospitalized SARS-CoV-2-infected subjects. Using trimeric spike glycoprotein, rather than nucleocapsid, enabled detection of responses in individuals with low antibody responses. IgG1 and IgG3 predominate to both antigens, but more anti-spike IgG1 than IgG3 was detectable. All antigens were effective for detecting responses in hospitalized patients. Anti-spike IgG, IgA and IgM antibody responses were readily detectable in saliva from a minority of RT-PCR confirmed, non-hospitalized symptomatic individuals, and these were mostly subjects who had the highest levels of anti-spike serum antibodies. Therefore, detecting antibody responses in both saliva and serum can contribute to determining virus exposure and understanding immune responses after SARS-CoV-2 infection.
中文翻译：
开发高灵敏度 ELISA 检测血清和唾液中 SARS-CoV-2 刺突糖蛋白的 IgG、IgA 和 IgM 抗体
检测 SARS-CoV-2 感染期间和感染后的抗体反应对于确定病毒的血清流行病学以及抗体在疾病中的潜在作用至关重要。可扩展、灵敏且特异的血清学检测对于这一过程至关重要。事实证明，重症住院患者的抗体检测相对简单；事实证明，检测患有轻度疾病和无症状感染者的反应不太可靠。我们假设抗体测定的次优灵敏度和抗体反应的区室化可能导致这种效应。我们系统地开发了一种 ELISA，优化了非住院 SARS-CoV-2 感染受试者的血清和唾液中的不同抗原和扩增步骤。使用三聚体刺突糖蛋白而不是核衣壳，能够检测抗体反应低的个体的反应。两种抗原均以 IgG1 和 IgG3 为主，但可检测到的抗尖峰 IgG1 多于 IgG3。所有抗原均可有效检测住院患者的反应。在少数 RT-PCR 确诊的非住院有症状个体的唾液中可以轻松检测到抗尖峰 IgG、IgA 和 IgM 抗体反应，并且这些人大多具有最高水平的抗尖峰血清抗体。因此，检测唾液和血清中的抗体反应有助于确定病毒暴露情况并了解 SARS-CoV-2 感染后的免疫反应。
更新日期：2021-05-01
中文翻译：
开发高灵敏度 ELISA 检测血清和唾液中 SARS-CoV-2 刺突糖蛋白的 IgG、IgA 和 IgM 抗体
检测 SARS-CoV-2 感染期间和感染后的抗体反应对于确定病毒的血清流行病学以及抗体在疾病中的潜在作用至关重要。可扩展、灵敏且特异的血清学检测对于这一过程至关重要。事实证明，重症住院患者的抗体检测相对简单；事实证明，检测患有轻度疾病和无症状感染者的反应不太可靠。我们假设抗体测定的次优灵敏度和抗体反应的区室化可能导致这种效应。我们系统地开发了一种 ELISA，优化了非住院 SARS-CoV-2 感染受试者的血清和唾液中的不同抗原和扩增步骤。使用三聚体刺突糖蛋白而不是核衣壳，能够检测抗体反应低的个体的反应。两种抗原均以 IgG1 和 IgG3 为主，但可检测到的抗尖峰 IgG1 多于 IgG3。所有抗原均可有效检测住院患者的反应。在少数 RT-PCR 确诊的非住院有症状个体的唾液中可以轻松检测到抗尖峰 IgG、IgA 和 IgM 抗体反应，并且这些人大多具有最高水平的抗尖峰血清抗体。因此，检测唾液和血清中的抗体反应有助于确定病毒暴露情况并了解 SARS-CoV-2 感染后的免疫反应。
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