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.
【저자키워드】 Biochemistry, Microbiology, 【초록키워드】 SARS-CoV-2, SARS-CoV-2 virus, RT-PCR, Antigen, sensitivity, specificity, Testing, Culture, lysis buffer, SARS-CoV-2 RNA, virus culture, real-time RT-PCR, nucleocapsid antigen, Simoa, real time, clinical sensitivity, 95% CI, house, SARS-CoV-2 infected patients, oropharyngeal swabs, cut-off, routine clinical practice, positive, reference method, SARS-CoV-2 nucleocapsid, ambition, analyzed, was used, Sample, develop, clinically, treated, can be used, demonstrated, correlated, comparable, Point, SARS-CoV-2 infected patient, 【제목키워드】 Antigen, oropharyngeal swab, SARS-CoV-2 nucleocapsid,