Serum SARS-COV-2 Nucleocapsid Protein: A Sensitivity and Specificity Early Diagnostic Marker for SARS-COV-2 Infection

Abstract

Objective: To explore the diagnostic value of serum severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid (N) protein assay in the early stages of SARS-COV-2 infection. Methods: Serum N protein level in SARS-COV-2 infected patients and non-SARS-COV-2 infected population was measured by enzyme-linked immunosorbent assay (ELISA) double antibody sandwich assay. Colloidal gold immunochromatography assay was used to detect serum N protein antibodies in the above populations. Results: Fifty cases of SARS-CoV-2 nucleic acid-positive and SARS-CoV-2 antibody-negative patients had a serum N protein positivity rate of 76%. Thirty-seven patients who were positive for serum SARS-CoV-2 antibody after infection had a serum SARS-CoV-2 N protein positivity rate of 2.7%. Serum N protein test results of 633 non-SARS-COV-2 infected patients, including pregnant women, patients with other respiratory infections, and individuals with increased rheumatoid factor were all negative, with serum N protein concentration <10.00 pg/mL at 100% specificity. Using SPSS 19.0 to calculate the receiver operating characteristic curve, the area under the curve was determined to be 0.9756 (95% confidence interval 0.9485-1.000, p < 0.0001), and sensitivity and specificity were 92% (95% confidence interval 81.16-96.85%) and 96.84% (95% confidence interval 95.17-97.15%), respectively. The best CUT-OFF value was 1.850 pg/mL. Conclusion: The measurement of serum SARS-COV-2 N protein has a high diagnostic value for infected patients before the antibody appears and shortens the window period of serological diagnosis. It is recommended that the manufacturer establish two different CUT-OFF values according to the purpose of the application. One CUT-OFF value is used for the diagnosis of clinical SARS-COV-2 infection, and the other is used to screen out as many suspected cases as possible.

Keywords: COVID-19; SARS-CoV-2; colloidal gold immunochromatography; diagnostic value; nucleocapsid protein.

Figure 1

CT images of 4 patients whose N protein is consistently negative, (A,B), CT images of Patient 3 1–3 days after admission; (C,D), CT images of Patient 32 1–3 days after admission; (E,F), CT images of Patient 33 1–3 days after admission; (G,H), CT images of Patient 40 1–3 days after admission.

Figure 2

ROC curve of serum SARS-COV-2 N protein, area under the ROC curve is 0.9756, 95% confidence interval (CI) is 0.9485 to 1.000, p-value is lower 0.0001, sensitivity and specificity are 92% (95% CI 81.16 to 96.85%) and 96.84% (95% CI 95.17 to 97.15%), respectively. The best CUT-OFF value is 1.850 pg/mL.

Figure 3

Distribution of N Protein concentration result of sample data using the CUT-OFF value equaling to 10, Group A (Nucleic Acid Positive + Antibody Negative, N = 50), Group B (Nucleic Acid Positive + Antibody Positive, N = 37), Control Group (non-SARS-COV-2 infection, N = 633).