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Evaluation of antibody responses against SARS coronaviral nucleocapsid or spike proteins by immunoblotting or ELISA.
J Med Virol. 2004 Jul; 73(3):338-46.JM

Abstract

Severe acute respiratory syndrome (SARS)-CoV is a newly emerging virus that causes SARS with high mortality rate in infected people. To study the humoral responses against SARS-CoV, we evaluated nucleocapsid (N) and spike (S) proteins-specific antibodies in patients' sera by Western blotting and enzyme-linked immunosorbent assay (ELISA). Recombinant N and S proteins of SARS-CoV were purified from transformed E. coli. Serum specimens from 40 SARS-CoV-infected patients in the convalescent phase were analyzed by Western blotting using the purified antigens. Serial serum specimens from 12 RT-PCR-confirmed SARS patients were assayed by ELISA using the recombinant N protein as coated antigen. By Western blotting, 97.5% of the SARS patients were positive for N protein-specific antibodies whereas only 47.5% of the samples were positive for S protein-specific antibodies. Using N protein-based ELISA, 10 out of the 12 patients were positive for N protein-specific antibodies and 6 of them showed seroconversion at mean of 16 days after onset of fever. Immunoblotting was useful for detecting the humoral immune response after SARS-CoV infection. Antibodies against SARS-CoV N protein appear at the early stage of infection, therefore, N protein-based ELISA could serve as a simple, sensitive, and specific test for diagnosing SARS-CoV infection.

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Authors+Show Affiliations

Huang LR
Graduate Institute of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan.
Chiu CM
No affiliation info available
Yeh SH
No affiliation info available
Huang WH
No affiliation info available
Hsueh PR
No affiliation info available
Yang WZ
No affiliation info available
Yang JY
No affiliation info available
Su IJ
No affiliation info available
Chang SC
No affiliation info available
Chen PJ
No affiliation info available

MeSH

Amino Acid SequenceAntibodies, ViralAntibody FormationAntigens, ViralBlotting, WesternCloning, MolecularCoronavirus Nucleocapsid ProteinsEnzyme-Linked Immunosorbent AssayEscherichia coliHumansMembrane GlycoproteinsMolecular Sequence DataNucleocapsid ProteinsRecombinant Fusion ProteinsSevere acute respiratory syndrome-related coronavirusSevere Acute Respiratory SyndromeSpike Glycoprotein, CoronavirusViral Envelope Proteins

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15170626

Citation

Huang, Li-Rung, et al. "Evaluation of Antibody Responses Against SARS Coronaviral Nucleocapsid or Spike Proteins By Immunoblotting or ELISA." Journal of Medical Virology, vol. 73, no. 3, 2004, pp. 338-46.
Huang LR, Chiu CM, Yeh SH, et al. Evaluation of antibody responses against SARS coronaviral nucleocapsid or spike proteins by immunoblotting or ELISA. J Med Virol. 2004;73(3):338-46.
Huang, L. R., Chiu, C. M., Yeh, S. H., Huang, W. H., Hsueh, P. R., Yang, W. Z., Yang, J. Y., Su, I. J., Chang, S. C., & Chen, P. J. (2004). Evaluation of antibody responses against SARS coronaviral nucleocapsid or spike proteins by immunoblotting or ELISA. Journal of Medical Virology, 73(3), 338-46.
Huang LR, et al. Evaluation of Antibody Responses Against SARS Coronaviral Nucleocapsid or Spike Proteins By Immunoblotting or ELISA. J Med Virol. 2004;73(3):338-46. PubMed PMID: 15170626.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Evaluation of antibody responses against SARS coronaviral nucleocapsid or spike proteins by immunoblotting or ELISA. AU - Huang,Li-Rung, AU - Chiu,Chi-Ming, AU - Yeh,Shiou-Hwei, AU - Huang,Wen-Hung, AU - Hsueh,Po-Ren, AU - Yang,Wen-Zieh, AU - Yang,Jyh-Yuan, AU - Su,Ih-Jen, AU - Chang,Shan-Chwen, AU - Chen,Pei-Jer, PY - 2004/6/2/pubmed PY - 2004/12/22/medline PY - 2004/6/2/entrez SP - 338 EP - 46 JF - Journal of medical virology JO - J Med Virol VL - 73 IS - 3 N2 - Severe acute respiratory syndrome (SARS)-CoV is a newly emerging virus that causes SARS with high mortality rate in infected people. To study the humoral responses against SARS-CoV, we evaluated nucleocapsid (N) and spike (S) proteins-specific antibodies in patients' sera by Western blotting and enzyme-linked immunosorbent assay (ELISA). Recombinant N and S proteins of SARS-CoV were purified from transformed E. coli. Serum specimens from 40 SARS-CoV-infected patients in the convalescent phase were analyzed by Western blotting using the purified antigens. Serial serum specimens from 12 RT-PCR-confirmed SARS patients were assayed by ELISA using the recombinant N protein as coated antigen. By Western blotting, 97.5% of the SARS patients were positive for N protein-specific antibodies whereas only 47.5% of the samples were positive for S protein-specific antibodies. Using N protein-based ELISA, 10 out of the 12 patients were positive for N protein-specific antibodies and 6 of them showed seroconversion at mean of 16 days after onset of fever. Immunoblotting was useful for detecting the humoral immune response after SARS-CoV infection. Antibodies against SARS-CoV N protein appear at the early stage of infection, therefore, N protein-based ELISA could serve as a simple, sensitive, and specific test for diagnosing SARS-CoV infection. SN - 0146-6615 UR - https://www.unboundmedicine.com/medline/citation/15170626/Evaluation_of_antibody_responses_against_SARS_coronaviral_nucleocapsid_or_spike_proteins_by_immunoblotting_or_ELISA_ DB - PRIME DP - Unbound Medicine ER -
Grapherence [↓48 ↑24]

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