Development and characterization of an indirect ELISA to detect SARS-CoV-2 spike protein-specific antibodies.
Development and characterization of an indirect ELISA to detect SARS-CoV-2 spike protein-specific antibodies.
复制标题
检测SARS-CoV-2刺突蛋白特异性抗体的间接酶联免疫吸附试验方法的建立及特性
DOI：
10.1016/j.jim.2021.112958
复制
发表时间：
2021-03
影响因子：
2.2
通讯作者：
Becker S
中科院分区：
文献类型：
--
作者：
Krähling V;Halwe S;Rohde C;Becker D;Berghöfer S;Dahlke C;Eickmann M;Ercanoglu MS;Gieselmann L;Herwig A;Kupke A;Müller H;Neubauer-Rädel P;Klein F;Keller C;Becker S
当前的严重急性呼吸综合征相关冠状病毒2型(SARS-CoV-2)大流行是一种国际关注的突发公共卫生事件。迫切需要灵敏和精确的诊断工具。在本研究中，我们建立了检测SARS-CoV-2特异性抗体的SARS-CoV-2刺突(S1)蛋白酶联免疫吸附试验(ELISA)。SARS-CoV-2和S1的特异性为97.8%(95%CI，96.7%-98.5%)，重复性好，精密度高(批内变异系数(CV)5.3%，批间变异系数(CV)7.9%)。标准曲线和每毫升任意酶联免疫吸附试验单位的内插有助于减少不同检测和操作人员之间的变异性。通过使用MERS冠状病毒和hCoV HKU1特异性抗体阳性的血清来解决与其他人类冠状病毒的交叉反应。监测2019年29例冠状病毒病(−-CoV-19)患者的23份单一样本的各种样本中的抗体发展情况，结果显示所有病例均有针对正宗SARS-CoV-2的血清转换和中和抗体。将SARS-CoV-2(S1)的酶联免疫吸附试验与市售的检测方法进行了比较，结果表明该方法具有较好的敏感性。结果表明，该方法具有较高的重复性、特异性和敏感性，适用于SARS-CoV-2和S1蛋白特异性抗体反应的检测。
The current Severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) pandemic is a public health emergency of international concern. Sensitive and precise diagnostic tools are urgently needed. In this study, we developed a SARS-CoV-2 spike (S1) protein enzyme-linked immunosorbent assay (ELISA) to detect SARS-CoV-2-specific antibodies. The SARS-CoV-2 S1 ELISA was found to be specific [97.8% (95% CI, 96.7% - 98.5%)], reproducible and precise (intra-assay coefficient of variability (CV) 5.3%, inter-assay CV 7.9%). A standard curve and the interpolation of arbitrary ELISA units per milliliter served to reduce the variability between different tests and operators. Cross-reactivity to other human coronaviruses was addressed by using sera positive for MERS-CoV- and hCoV HKU1-specific antibodies. Monitoring antibody development in various samples of twenty-three and single samples of twenty-nine coronavirus disease 2019 (COVID−19) patients revealed seroconversion and neutralizing antibodies against authentic SARS-CoV-2 in all cases. The comparison of the SARS-CoV-2 (S1) ELISA with a commercially available assay showed a better sensitivity for the in-house ELISA. The results demonstrate a high reproducibility, specificity and sensitivity of the newly developed ELISA, which is suitable for the detection of SARS-CoV-2 S1 protein-specific antibody responses.
登录
查看更多内容
影响因子：
--
作者：
Hulswit RJ;de Haan CA;Bosch BJ
通讯作者：
Bosch BJ
影响因子：
13.2
作者：
Sun, Baoqing;Feng, Ying;Chen, Ling
通讯作者：
Chen, Ling
DOI：
10.1016/j.jcv.2020.104394
发表时间：
2020-07-01
期刊：
Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology
影响因子：
--
作者：
Kruttgen, Alexander;Cornelissen, Christian G;Kleines, Michael
通讯作者：
Kleines, Michael
影响因子：
12.7
作者：
Kohmer, Niko;Westhaus, Sandra;Rabenau, Holger F.
通讯作者：
Rabenau, Holger F.
影响因子：
10.7
作者：
Shereen, Muhammad Adnan;Khan, Suliman;Siddique, Rabeea
通讯作者：
Siddique, Rabeea