Relative rates of non-pneumonic SARS coronavirus infection and SARS coronavirus pneumonia

Abstract

Background: Although the genome of severe acute respiratory syndrome coronavirus (SARS-CoV) has been sequenced and a possible animal reservoir identified, seroprevalence studies and mass screening for detection of subclinical and non-pneumonic infections are still lacking.

Methods: We cloned and purified the nucleocapsid protein and spike polypeptide of SARS-CoV and examined their immunogenicity with serum from patients with SARS-CoV pneumonia. An ELISA based on recombinant nucleocapsid protein for IgG detection was tested with serum from 149 healthy blood donors who donated 3 years previously and with serum positive for antibodies against SARS-CoV (by indirect immunofluorescence assay) from 106 patients with SARS-CoV pneumonia. The seroprevalence of SARS-CoV was studied with the ELISA in healthy blood donors who donated during the SARS outbreak in Hong Kong, non-pneumonic hospital inpatients, and symptom-free health-care workers. All positive samples were confirmed by two separate western-blot assays (with recombinant nucleocapsid protein and recombinant spike polypeptide).

Findings: Western-blot analysis showed that the nucleocapsid protein and spike polypeptide of SARS-CoV are highly immunogenic. The specificity of the IgG antibody test (ELISA with positive samples confirmed by the two western-blot assays) was 100%, and the sensitivity was 94.3%. Three of 400 healthy blood donors who donated during the SARS outbreak and one of 131 non-pneumonic paediatric inpatients were positive for IgG antibodies, confirmed by the two western-blot assays (total, 0.48% of our study population).

Interpretation: Our findings support the existence of subclinical or non-pneumonic SARS-CoV infections. Such infections are more common than SARS-CoV pneumonia in our locality.

Figure 1

Western-blot analysis of purified recombinant SARS-CoV nucleocapsid protein and purified recombinant SARS-CoV spike polypeptide Prominent immunoreactive protein bands of about 50kDa were visible for both antigens with serum from three patients with SARS-CoV pneumonia, indicating antigen–antibody interactions between the recombinant SARS-CoV nucleocapsid protein and the patients'antibodies (lanes 1–3). No immunoreactive band was detected for serum from three healthy blood donors (lanes 4–6).

Figure 2

ELISA based on recombinant nucleocapsid protein IgG antibody for SARS-CoV infection