Development and Evaluation of Quantitative Immunoglobulin G Enzyme-Linked Immunosorbent Assay for the Diagnosis of Coronavirus Disease 2019 Using Truncated Recombinant Nucleocapsid Protein as Assay Antigen.
Development and Evaluation of Quantitative Immunoglobulin G Enzyme-Linked Immunosorbent Assay for the Diagnosis of Coronavirus Disease 2019 Using Truncated Recombinant Nucleocapsid Protein as Assay Antigen.
复制标题
使用截短重组核衣壳蛋白作为检测抗原的定量免疫球蛋白G酶联免疫吸附试验2019用于冠状病毒病诊断的开发和评价。
DOI：
10.3390/ijerph18189630
复制
发表时间：
2021-09-13
影响因子：
--
通讯作者：
Morita K
中科院分区：
文献类型：
--
作者：
Mutantu PN;Ngwe Tun MM;Nabeshima T;Yu F;Mukadi PK;Tanaka T;Tashiro M;Fujita A;Kanie N;Oshiro R;Takazono T;Imamura Y;Hirayama T;Moi ML;Inoue S;Izumikawa K;Yasuda J;Morita K
严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)是冠状病毒病2019年(新冠肺炎)的病原体。实时荧光定量RT-PCR法是目前诊断新冠肺炎最常用的方法。然而，作为RT-PCR的补充工具，迫切需要血清学检测。Hachim等人。2020和Burbelo等人。2020年发现，抗核衣壳蛋白(N)SARS-CoV-2抗体高于尖峰抗体且出现时间较早。此外，针对N蛋白的交叉反应抗体比针对Spike蛋白的抗体更普遍。以截短的重组SARS冠状病毒N蛋白为检测抗原，建立了一种低交叉反应的免疫球蛋白G(Ig G)间接ELISA。删除冠状病毒N蛋白的一个高度保守区，并利用大肠杆菌蛋白表达系统制备该蛋白。以空斑减少中和试验和商品化的SARS-CoV-2刺突蛋白免疫吸附试验为金标准，对从新冠肺炎疑似病例中采集的177份标本和新冠肺炎前期间采集的155份阴性对照血清进行了检测。SARS-CoV-2N截短蛋白酶联免疫吸附试验的敏感性(91.1%vs.91.9%)和特异性(93.8%vs.93.8%)与SPEKE-Ig G(93.8%vs.93.8%)相似，而特异性(93.8%vs.89.9%)高于全长N蛋白(93.8%vs.89.9%)。本方法可用于新冠肺炎的诊断和监测。
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19). Real-time RT-PCR is the most commonly used method for COVID-19 diagnosis. However, serological assays are urgently needed as complementary tools to RT-PCR. Hachim et al. 2020 and Burbelo et al. 2020 demonstrated that anti-nucleocapsid(N) SARS-CoV-2 antibodies are higher and appear earlier than the spike antibodies. Additionally, cross-reactive antibodies against N protein are more prevalent than those against spike protein. We developed a less cross-reactive immunoglobulin G (IgG) indirect ELISA by using a truncated recombinant SARS-CoV-2 N protein as assay antigen. A highly conserved region of coronaviruses N protein was deleted and the protein was prepared using an E. coli protein expression system. A total of 177 samples collected from COVID-19 suspected cases and 155 negative control sera collected during the pre-COVID-19 period were applied to evaluate the assay’s performance, with the plaque reduction neutralization test and the commercial SARS-CoV-2 spike protein IgG ELISA as gold standards. The SARS-CoV-2 N truncated protein-based ELISA showed similar sensitivity (91.1% vs. 91.9%) and specificity (93.8% vs. 93.8%) between the PRNT and spike IgG ELISA, as well as also higher specificity compared to the full-length N protein (93.8% vs. 89.9%). Our ELISA can be used for the diagnosis and surveillance of COVID-19.
登录
查看更多内容
影响因子：
2.2
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
通讯作者：
Becker S
影响因子：
9.4
作者：
Jiao, Yongjun;Zeng, Xiaoyan;Wang, Hua
通讯作者：
Wang, Hua
影响因子：
2.2
作者：
Shirato, Kazuya;Tomita, Yuriko;Takeda, Makoto
通讯作者：
Takeda, Makoto
影响因子：
30.5
作者：
Hachim, Asmaa;Kavian, Niloufar;Valkenburg, Sophie A.
通讯作者：
Valkenburg, Sophie A.
影响因子：
3.1
作者：
He Q;Du Q;Lau S;Manopo I;Lu L;Chan SW;Fenner BJ;Kwang J
通讯作者：
Kwang J