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Comparative Study
. 2005 Jul;43(7):3054-8.
doi: 10.1128/JCM.43.7.3054-3058.2005.

Differential sensitivities of severe acute respiratory syndrome (SARS) coronavirus spike polypeptide enzyme-linked immunosorbent assay (ELISA) and SARS coronavirus nucleocapsid protein ELISA for serodiagnosis of SARS coronavirus pneumonia

Patrick C Y Woo  1 , Susanna K P Lau, Beatrice H L Wong, Hoi-Wah Tsoi, Ami M Y Fung, Richard Y T Kao, Kwok-Hung Chan, J S Malik Peiris, Kwok-Yung Yuen
Affiliations
Comparative Study

Differential sensitivities of severe acute respiratory syndrome (SARS) coronavirus spike polypeptide enzyme-linked immunosorbent assay (ELISA) and SARS coronavirus nucleocapsid protein ELISA for serodiagnosis of SARS coronavirus pneumonia

Patrick C Y Woo et al. J Clin Microbiol. 2005 Jul.

Abstract

The use of recombinant severe acute respiratory syndrome-coronavirus (SARS-CoV) nucleocapsid protein (N) enzyme-linked immunosorbent assay (ELISA)-based antibody and antigen tests for diagnosis of SARS-CoV infections have been widely reported. However, no recombinant SARS-CoV spike protein (S)-based ELISA is currently available. In this article, we describe the problems and solutions of setting up the recombinant SARS-CoV S-based ELISA for antibody detection. The SARS-CoV S-based immunoglobulin M (IgM) and IgG ELISAs were evaluated and compared with the corresponding N-based ELISA for serodiagnosis of SARS-CoV pneumonia, using sera from 148 healthy blood donors who donated blood 3 years ago as controls and 95 SARS-CoV pneumonia patients in Hong Kong. Results obtained by the recombinant S (rS)-based IgG ELISA using the regenerated S prepared by dialysis with decreasing concentrations of urea or direct addition of different coating buffers, followed by addition of different regeneration buffer, identified 4 M urea and 1 M sarcosine for plate coating and no regeneration buffer as the most optimal conditions for antibody detection. The specificities of the S-based ELISA for IgG and IgM detection were 98.6% and 93.9%, with corresponding sensitivities of 58.9% and 74.7%, respectively. The sensitivity of the rN IgG ELISA (94.7%) is significantly higher than that of the rS IgG ELISA (P < 0.001), whereas the sensitivity of the rS IgM ELISA is significantly higher than that of the rN IgM ELISA (55.2%) (P < 0.01). An ELISA for detection of IgM against S and N could be more sensitive than one that detects IgM against N alone for serodiagnosis of SARS-CoV pneumonia.

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Figures

FIG. 1.
FIG. 1.
Evaluation of sensitivity and specificity of rS-based IgG (A) and IgM (B) antibody ELISA for SARS-CoV pneumonia. Serum specimens were obtained from 95 patients with SARS-CoV pneumonia, and control serum specimens were obtained from 148 healthy blood donors. The test results were plotted as OD450 values. The cutoff line for positive diagnosis is drawn at a value that equals the sum of the mean value and two times the standard deviation for the healthy blood donors.
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References

    1. Briese, T., C. G. Hatalski, S. Kliche, Y. S. Park, and W. I. Lipkin. 1995. Enzyme-linked immunosorbent assay for detecting antibodies to Borne disease virus-specific proteins. J. Clin. Microbiol. 33:348-351. - PMC - PubMed
    1. Chan, C. M., P. C. Y. Woo, A. S. P. Leung, S. K. P. Lau, X. Y. Che, L. Cao, and K. Y. Yuen. 2002. Detection of specific antibodies to an antigenic cell wall galactomannoprotein for serodiagnosis of Aspergillus fumigatus aspergillosis. J. Clin. Microbiol. 40:2041-2045. - PMC - PubMed
    1. Chan, P. K., W. K. To, E. Y. Liu, T. K. Ng, J. S. Tam, J. J. Sung, J. M. Lacroix, and M. Houde. 2004. Evaluation of a peptide-based enzyme immunoassay for anti-SARS coronavirus IgG antibody. J. Med. Virol. 74:517-520. - PMC - PubMed
    1. Che, X. Y., L.W. Qiu, Y. X. Pan, K. Wen, W. Hao, L. Y. Zhang, Y. D. Wang, Z. Y. Liao, X. Hua, V. C. Cheng, and K. Y. Yuen. 2004. Sensitive and specific monoclonal antibody-based capture enzyme immunoassay for detection of nucleocapsid antigen in sera from patients with severe acute respiratory syndrome. J. Clin. Microbiol. 42:2629-2635. - PMC - PubMed
    1. Drosten, C., S. Günther, W. Preiser, S. van der Werf, H. R. Brodt, S. Becker, H. Rabenau, M. Panning, L. Kolesnikova, R. A. M. Fouchier, A. Berger, A. M. Burguière, J. Cinatl, M. Eickmann, N. Escriou, K. Grywna, S. Kramme, J. C. Manuguerra, S. Müller, V. Rickerts, M. Stürmer, S. Vieth, H. D. Klenk, A. D. M. E. Osterhaus, H. Schmitz, and H. W. Doerr. 2003. Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N. Engl. J. Med. 348:1967-1976. - PubMed

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