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Immunological Characterization of the Spike Protein of the Severe Acute Respiratory Syndrome Coronavirus

Animal Health Biotechnology Unit, Temasek Life Science Laboratory, National University of Singapore,¹ Department of Rheumatology, Allergy, and Immunology, Tan Tock Seng Hospital,² Virology Section, Department of Pathology, Singapore General Hospital,³ Environment Health Institute, National Environment Agency, Singapore⁴

Received 24 November 2003/ Returned for modification 1 January 2004/ Accepted 7 January 2004

Severe acute respiratory syndrome (SARS) is a novel infectious disease caused by the SARS-associated coronavirus (SARS-CoV). There are four major structural proteins in the SARS-CoV, including the nucleocapsid, spike, membrane, and small envelope proteins. In this study, two sets of truncated fragments of spike protein were generated, the first were approximately 210-bp nonoverlapping fragments and the second were overlapping segments of 750 to 900 bp. From these 23 fragments, we identified a fragment of 259 amino acids (amino acids 441 to 700) that is a major immunodominant epitope. This fragment was highly expressed, and the purified fragment C could detect all 33 SARS patient serum samples tested, collected from 7 to 60 days after the onset of fever, but had no reactivity with all 66 healthy human serum samples tested. Thus, fragment C of spike protein was identified as an immunodominant antigen and could be used for serological detection of SARS-CoV infection.

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