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Clinical and Diagnostic Laboratory Immunology, March 2004, p. 362-371, Vol. 11, No. 2
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.2.362-371.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Profiles of Antibody Responses against Severe Acute Respiratory Syndrome Coronavirus Recombinant Proteins and Their Potential Use as Diagnostic Markers

Yee-Joo Tan,1,{dagger}* Phuay-Yee Goh,1,{dagger} Burtram C. Fielding,1 Shuo Shen,1 Chih-Fong Chou,1 Jian-Lin Fu,1 Hoe Nam Leong,2 Yee Sin Leo,2 Eng Eong Ooi,3 Ai Ee Ling,4 Seng Gee Lim,1,{ddagger} and Wanjin Hong1,{ddagger}

Institute of Molecular and Cell Biology,1 Tan Tock Seng Hospital,2 Environmental Health Institute, National Environmental Agency,3 Virology Section, Department of Pathology, Singapore General Hospital,Singapore, Republic of Singapore4

Received 29 September 2003/ Returned for modification 24 November 2003/ Accepted 22 December 2003

A new coronavirus (severe acute respiratory syndrome coronavirus [SARS-CoV]) has been identified to be the etiological agent of severe acute respiratory syndrome. Given the highly contagious and acute nature of the disease, there is an urgent need for the development of diagnostic assays that can detect SARS-CoV infection. For determination of which of the viral proteins encoded by the SARS-CoV genome may be exploited as diagnostic antigens for serological assays, the viral proteins were expressed individually in mammalian and/or bacterial cells and tested for reactivity with sera from SARS-CoV-infected patients by Western blot analysis. A total of 81 sera, including 67 from convalescent patients and seven pairs from two time points of infection, were analyzed, and all showed immunoreactivity towards the nucleocapsid protein (N). Sera from some of the patients also showed immunoreactivity to U274 (59 of 81 [73%]), a protein that is unique to SARS-CoV. In addition, all of the convalescent-phase sera showed immunoreactivity to the spike (S) protein when analyzed by an immunofluorescence method utilizing mammalian cells stably expressing S. However, samples from the acute phase (2 to 9 days after the onset of illness) did not react with S, suggesting that antibodies to N may appear earlier than antibodies to S. Alternatively, this could be due to the difference in the sensitivities of the two methods. The immunoreactivities to these recombinant viral proteins are highly specific, as sera from 100 healthy donors did not react with any of them. These results suggest that recombinant N, S, and U274 proteins may be used as antigens for the development of serological assays for SARS-CoV.

* Corresponding author. Mailing address: Institute of Molecular and Cell Biology, 30 Medical Dr., Singapore 117609, Republic of Singapore. Phone: 65-68743780. Fax: 65-67791117. E-mail: mcbtanyj{at}imcb.nus.edu.sg.

{dagger} Y.-J.T. and P.-Y.G. contributed equally to this study.

S.G.L. and W.H. were both senior authors for this paper.

Clinical and Diagnostic Laboratory Immunology, March 2004, p. 362-371, Vol. 11, No. 2
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.2.362-371.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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