Development of a high-sensitivity ELISA detecting IgG, IgA and IgM antibodies to the SARS-CoV-2 spike glycoprotein in serum and saliva.
Development of a high-sensitivity ELISA detecting IgG, IgA and IgM antibodies to the SARS-CoV-2 spike glycoprotein in serum and saliva.
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血清和唾液SARS-CoV-2刺突糖蛋白IgG、伊加和IgM抗体高灵敏度ELISA检测方法的建立
DOI：
10.1111/imm.13349
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发表时间：
2021-09
期刊：
影响因子：
6.4
通讯作者：
Richter AG
中科院分区：
文献类型：
--
作者：
Faustini SE;Jossi SE;Perez-Toledo M;Shields AM;Allen JD;Watanabe Y;Newby ML;Cook A;Willcox CR;Salim M;Goodall M;Heaney JL;Marcial-Juarez E;Morley GL;Torlinska B;Wraith DC;Veenith TV;Harding S;Jolles S;Ponsford MJ;Plant T;Huissoon A;O'Shea MK;Willcox BE;Drayson MT;Crispin M;Cunningham AF;Richter AG
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，很容易检测到抗尖峰 IgG、IgA 和 IgM 抗体反应，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。本手稿描述了一种高灵敏度 ELISA 的开发，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，可以检测轻度至中度感染个体的反应。血清和唾液中抗体的检测有助于确定病毒暴露情况并了解对 SARS-CoV-2 的免疫反应。
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. This manuscript describes the development of a highly sensitive ELISA, optimizing different antigens and amplification steps, in serum and saliva from non‐hospitalized SARS‐CoV‐2‐infected subjects. Using a trimeric spike glycoprotein, rather than nucleocapsid, enabled detection of responses in individuals with mild‐to‐moderate infection. The detection of antibodies in both serum and saliva can contribute to determining virus exposure and understanding immune responses to SARS‐CoV‐2.
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影响因子：
5.4
作者：
Lau, Susanna K. P.;Lee, Paul;Yuen, Kwok-Yung
通讯作者：
Yuen, Kwok-Yung
影响因子：
13.2
作者：
Sun, Baoqing;Feng, Ying;Chen, Ling
通讯作者：
Chen, Ling
影响因子：
2.2
作者：
Mestecky, J;Hamilton, RG;Reimer, CB
通讯作者：
Reimer, CB
影响因子：
24.8
作者：
Isho B;Abe KT;Zuo M;Jamal AJ;Rathod B;Wang JH;Li Z;Chao G;Rojas OL;Bang YM;Pu A;Christie-Holmes N;Gervais C;Ceccarelli D;Samavarchi-Tehrani P;Guvenc F;Budylowski P;Li A;Paterson A;Yue FY;Marin LM;Caldwell L;Wrana JL;Colwill K;Sicheri F;Mubareka S;Gray-Owen SD;Drews SJ;Siqueira WL;Barrios-Rodiles M;Ostrowski M;Rini JM;Durocher Y;McGeer AJ;Gommerman JL;Gingras AC
通讯作者：
Gingras AC
DOI：
10.1093/infdis/jiaa593
发表时间：
2020-11-13
期刊：
The Journal of infectious diseases
影响因子：
--
作者：
Schwaiger J;Karbiener M;Aberham C;Farcet MR;Kreil TR
通讯作者：
Kreil TR