评估 SARS-CoV-2 捕获 IgM 抗体检测试剂盒在恢复期血清中的应用。
Evaluation of a SARS-CoV-2 Capture IgM Antibody Assay in Convalescent Sera.
机构信息
Division of Pediatric Infectious Diseases, Emory University School of Medicinegrid.471395.d and Children's Health Care of Atlanta, Atlanta, Georgia, USA.
Division of Infectious Diseases, Department of Medicine, Emory University School of Medicinegrid.471395.d, Atlanta, Georgia, USA.
出版信息
Microbiol Spectr. 2021 Oct 31;9(2):e0045821. doi: 10.1128/Spectrum.00458-21. Epub 2021 Sep 8.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for a global pandemic with over 152 million cases and 3.19 million deaths reported by early May 2021. Understanding the serological response to SARS-CoV-2 is critical to determining the burden of infection and disease (coronavirus disease 2019 [COVID-19]) and transmission dynamics. We developed a capture IgM assay because it should have better sensitivity and specificity than the commonly used indirect assay. Here, we report the development and performance of a capture IgM enzyme-linked immunosorbent assay (ELISA) and a companion indirect IgG ELISA for the spike (S) and nucleocapsid (N) proteins and the receptor-binding domain (RBD) of S. We found that among the IgM ELISAs, the S ELISA was positive in 76% of 55 serum samples from SARS-CoV-2 PCR-positive patients, the RBD ELISA was positive in 55% of samples, and the N ELISA was positive in 15% of samples. The companion indirect IgG ELISAs were positive for S in 89% of the 55 serum samples, RBD in 78%, and N in 85%. While the specificities for IgM RBD, S, and N ELISAs and IgG S and RBD ELISAs were 97% to 100%, the specificity of the N IgG ELISA was lower (89%). RBD-specific IgM antibodies became undetectable by 3 to 6 months, and S IgM reached low levels at 6 months. The corresponding IgG S, RBD, and N antibodies persisted with some decreases in levels over this time period. These capture IgM ELISAs and the companion indirect IgG ELISAs should enhance serologic studies of SARS-CoV-2 infections. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has inflicted tremendous loss of lives, overwhelmed health care systems, and disrupted all aspects of life worldwide since its emergence in Wuhan, China, in December 2019. Detecting current and past infection by PCR or serology is important to understanding and controlling SARS-CoV-2. With increasing prevalence of past infection or vaccination, IgG antibodies are less helpful in diagnosing a current infection. IgM antibodies indicate a more recent infection and can supplement PCR diagnosis. We report an alternative method, capture IgM, to detect serum IgM antibodies, which should be more sensitive and specific than most currently used methods. We describe this capture IgM assay and a companion indirect IgG assay for the SARS-CoV-2 spike (S), nucleocapsid (N), and receptor-binding domain (RBD) proteins. These assays can add value to diagnostic and serologic studies of coronavirus disease 2019 (COVID-19).
严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)引发了一场全球大流行,截至 2021 年 5 月初,已报告超过 1.52 亿例病例和 319 万人死亡。了解对 SARS-CoV-2 的血清学反应对于确定感染和疾病(2019 年冠状病毒病[COVID-19])的负担以及传播动态至关重要。我们开发了一种捕获 IgM 测定法,因为它应该比常用的间接测定法具有更好的敏感性和特异性。在这里,我们报告了一种用于 Spike(S)和核衣壳(N)蛋白以及 S 的受体结合域(RBD)的捕获 IgM 酶联免疫吸附测定(ELISA)和配套的间接 IgG ELISA 的开发和性能。我们发现,在 55 份 SARS-CoV-2 PCR 阳性患者的血清样本中,IgM ELISA 中 76%的 S ELISA 呈阳性,RBD ELISA 呈阳性的占 55%,N ELISA 呈阳性的占 15%。配套的间接 IgG ELISA 对 55 份血清样本中的 S 呈阳性,对 RBD 呈阳性的占 78%,对 N 呈阳性的占 85%。IgM RBD、S 和 N ELISA 以及 IgG S 和 RBD ELISA 的特异性为 97%至 100%,而 N IgG ELISA 的特异性较低(89%)。RBD 特异性 IgM 抗体在 3 至 6 个月内无法检测到,而 6 个月时 S IgM 达到低水平。在此期间,相应的 IgG S、RBD 和 N 抗体的水平有所下降。这些捕获的 IgM ELISA 和配套的间接 IgG ELISA 应增强对 SARS-CoV-2 感染的血清学研究。自 2019 年 12 月在中国武汉出现以来,严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)已造成巨大的生命损失,使医疗保健系统不堪重负,并在全球范围内扰乱了生活的方方面面。通过 PCR 或血清学检测来检测当前和过去的感染非常重要,这有助于了解和控制 SARS-CoV-2。随着过去感染或接种疫苗的流行率增加,IgG 抗体在诊断当前感染方面的帮助较小。IgM 抗体表明最近发生了感染,并且可以补充 PCR 诊断。我们报告了一种替代方法,即捕获 IgM,用于检测血清 IgM 抗体,该方法应该比目前大多数常用方法更敏感和更特异。我们描述了这种用于 SARS-CoV-2 刺突(S),核衣壳(N)和受体结合域(RBD)蛋白的捕获 IgM 测定法和配套的间接 IgG 测定法。这些测定法可以为 2019 年冠状病毒病(COVID-19)的诊断和血清学研究增加价值。