Department of Biochemistry and Immunology, Lillebaelt Hospital, University Hospital of Southern Denmark, Vejle, Denmark.
Department of Regional Health Research, University of Southern Denmark, Odense, Denmark.
Sci Rep. 2021 Oct 13;11(1):20323. doi: 10.1038/s41598-021-99807-7.
This study aimed to develop a highly sensitive SARS-CoV-2 nucleocapsid antigen assay using the single molecule array (Simoa) technology and compare it with real time RT-PCR as used in routine clinical practice with the ambition to achieve a comparative technical and clinical sensitivity. Samples were available from 148 SARS-CoV-2 real time RT-PCR positive and 73 SARS-CoV-2 real time RT-PCR negative oropharyngeal swabs. For determination of technical sensitivity SARS-CoV-2 virus culture material was used. The samples were treated with lysis buffer and analyzed using both an in-house and a pre-commercial SARS-CoV-2 nucleocapsid antigen assay on Simoa. Both nucleocapsid antigen assays have a technical sensitivity corresponding to around 100 SARS-CoV-2 RNA molecules/mL. Using a cut-off at 0.1 pg/mL the pre-commercial SARS-CoV-2 nucleocapsid antigen assay had a sensitivity of 96% (95% CI 91.4-98.5%) and specificity of 100% (95% CI 95.1-100%). In comparison the in-house nucleocapsid antigen assay had sensitivity of 95% (95% CI 89.3-98.1%) and a specificity of 100% (95% CI 95.1-100%) using a cut-off at 0.01 pg/mL. The two SARS-CoV-2 nucleocapsid antigen assays correlated with r = 0.91 (P < 0.0001). The in-house and the pre-commercial SARS-CoV-2 nucleocapsid antigen assay demonstrated technical and clinical sensitivity comparable to real-time RT-PCR methods for identifying SARS-CoV-2 infected patients and thus can be used clinically as well as serve as a reference method for antigen Point of Care Testing.
本研究旨在开发一种基于单分子阵列（Simoa）技术的高灵敏度 SARS-CoV-2 核衣壳抗原检测方法，并与常规临床实践中使用的实时 RT-PCR 进行比较，旨在实现技术和临床灵敏度的可比性。 样品来自 148 份 SARS-CoV-2 实时 RT-PCR 阳性和 73 份 SARS-CoV-2 实时 RT-PCR 阴性或咽拭子。为了确定技术灵敏度，使用了 SARS-CoV-2 病毒培养物。 将样品用裂解缓冲液处理，然后使用基于 Simoa 的内部和预商业化 SARS-CoV-2 核衣壳抗原检测方法进行分析。 两种核衣壳抗原检测方法的技术灵敏度均对应约 100 SARS-CoV-2 RNA 分子/mL。 使用 0.1 pg/mL 作为截断值，预商业化的 SARS-CoV-2 核衣壳抗原检测方法的灵敏度为 96%（95%CI 91.4-98.5%），特异性为 100%（95%CI 95.1-100%）。 相比之下，使用 0.01 pg/mL 作为截断值，内部核衣壳抗原检测方法的灵敏度为 95%（95%CI 89.3-98.1%），特异性为 100%（95%CI 95.1-100%）。 两种 SARS-CoV-2 核衣壳抗原检测方法的相关性为 r=0.91（P<0.0001）。 内部和预商业化的 SARS-CoV-2 核衣壳抗原检测方法在识别 SARS-CoV-2 感染患者方面表现出与实时 RT-PCR 方法相当的技术和临床灵敏度，因此可在临床上使用，并可作为抗原即时检测的参考方法。
