Dataset Information

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

ABSTRACT:

Background

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.

Objectives

To develop and evaluate an ELISA system for detection of anti-S Abs in patient sera.

Study design

Express recombinant S450-650 in E. Coli and evaluate the sensitivity and specificity of an ELISA system based on the S450-650 polypeptide.

Results

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.

Conclusion

The S450-650-based ELISA can detect anti-S IgG Abs with high sensitivity and specificity.

SUBMITTER: Zhao J

PROVIDER: S-EPMC7108335 | biostudies-literature | 2005 May

REPOSITORIES: biostudies-literature

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Publications

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

Zhao Jincun J Wang Wei W Wang Guang-Fa GF Li Yonghua Y Zhuang Hui H Xu Xiaoyuan X Ren Furong F Zhao Zhendong Z Gao Xiao-Ming XM

Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology 20050501 1

<h4>Background</h4>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.<h4>Objectives</h4>To develop and evaluate an ELISA system for detection of anti-S Abs in patient sera.<h4>Study design</h4>Express recombinant S450-650 in E. Coli and evaluate the sensitivity and specificity of an ELISA ...[more]

PMID: 15797360

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Project description:Porcine deltacoronavirus (PDCoV) is an emerging porcine enteric coronavirus causing significant economic losses to the pig farming industry globally. In this study, we expressed the S protein of a highly virulent PDCoV strain in the CHO eukaryotic expression system. After immunizing alpaca with the PDCoV S protein and employing the phage display library technique, a high-affinity and specific nanobody Nb3 against PDCoV S protein was successfully established by three rounds of biopanning and a phage enzyme-linked immunosorbent assay (ELISA). Furthermore, a competitive ELISA (cELISA) was developed based on Nb3 to rapidly and efficiently detect PDCoV antibody levels. The cELISA displayed no cross-reaction with positive sera of porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine rotavirus (PoRV), pseudorabies virus (PRV), classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), or porcine circovirus 2 (PCV2), thereby showing good specificity. The cELISA successfully detected positive sera diluted 1:127 (percentage inhibition ≥ 50.02%), indicating high sensitivity. Both the intra- and inter-batch coefficients of variation were less than 10%, indicating good repeatability. The cELISA had a total coincidence rate of 98.33% with the indirect immunofluorescence assay and a significant positive correlation with the virus neutralization test (r = 0.861, P < 0.001), suggesting that the cELISA can be used to measure the neutralizing antibody titers in serum samples. In conclusion, our nanobody-based cELISA showed good performance indicators and can be used to monitor and evaluate antibody levels following clinical infection of PDCoV or vaccine immunization.ImportanceThis study screened out a high-affinity and specific nanobody Nb3 against porcine deltacoronavirus (PDCoV) S protein and established a nanobody-based competitive ELISA (cELISA) for PDCoV antibody detection. This cELISA is a simple, rapid, and specific method that can effectively measure the neutralizing antibody titers in serum samples.

Dataset Information

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

Background

Objectives

Study design

Results

Conclusion

Publications

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

Similar Datasets

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