Development and evaluation of an enzyme-linked immunosorbent assay for detection of antibodies against the spike protein of SARS-coronavirus

• Published in: Journal of clinical virology Vol. 33; no. 1; pp. 12 - 18
• Main Authors: Zhao, Jincun, Wang, Wei, Wang, Guang-Fa, Li, Yonghua, Zhuang, Hui, Xu, Xiaoyuan, Ren, Furong, Zhao, Zhendong, Gao, Xiao-Ming
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.05.2005; Elsevier Science; Published by Elsevier B.V
• Subjects: Adolescent; Adult; Antibodies; Antibodies, Viral - blood; Biological and medical sciences; ELISA; Enzyme-Linked Immunosorbent Assay; Female; Fundamental and applied biological sciences. Psychology; Human viral diseases; Humans; Immunoglobulin G - blood; Infectious diseases; Male; Medical sciences; Membrane Glycoproteins - genetics; Membrane Glycoproteins - immunology; Microbiology; Middle Aged; Miscellaneous; Recombinant Proteins - genetics; Recombinant Proteins - immunology; SARS coronavirus; SARS Virus; Sensitivity and Specificity; Severe Acute Respiratory Syndrome; Spike glycoprotein; Spike Glycoprotein, Coronavirus; Viral diseases; Viral Envelope Proteins - genetics; Viral Envelope Proteins - immunology; Virology; Antibodies; SARS-coronavirus; Spike glycoprotein; ELISA; Microbiology; Antibody; Glycoprotein; Protein; Virology; Virus; Coronavirus; Severe acute respiratory syndrome virus; Coronaviridae; Detection; ELISA assay; Nidovirales
• ISSN: 1386-6532; 1873-5967
• DOI: 10.1016/j.jcv.2004.09.024

Severe acute respiratory syndrome (SARS) is caused by infection with SARS-associated coronavirus (CoV). Amino acid residues 450–650 of the spike (S) glycoprotein of SARS-CoV (S450-650) contains dominant epitopes for anti-viral antibodies (Abs) in patient sera. To develop and evaluate an ELISA system for detection of anti-S Abs in patient sera. Express recombinant S450-650 in E. Coli and evaluate the sensitivity and specificity of an ELISA system based on the S450-650 polypeptide. The S450-650-based ELISA detected IgG Abs in 41 out of 51 serum samples from 22 hospitalized patients with probable SARS, a result closely correlated with that obtained with a virus-based ELISA ( r = 0.75, k = 0.8). Differential anti-S IgG responses were observed amongst SARS patients. Some of them produced anti-S Abs early during their infection, while others failed to make IgG Abs against the S450-650 polypeptide. None of the serum samples from 100 healthy blood donors was positive in the S450-650-based assay. The S450-650-based ELISA can detect anti-S IgG Abs with high sensitivity and specificity.