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Screening of specific antigens for SARS clinical diagnosis using a protein microarray
Dan-Dan Lu1, Su-Hong Chen, Shi-Meng Zhang
1Beijing Institute of Radiation Medicine, Beijing, 100850, People's Republic of China.
The Analyst
|March 19, 2005
Summary
A novel diagnostic antigen, GST-N2, was identified for severe acute respiratory syndrome coronavirus (SARS-CoV) antibody testing. This protein microarray-based method shows high sensitivity and specificity for detecting SARS antibodies in patient sera.
Area of Science:
- Virology
- Immunology
- Biotechnology
Background:
- Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) poses a significant public health threat.
- Accurate and sensitive diagnostic tools are crucial for managing SARS outbreaks.
Purpose of the Study:
- To identify and validate a highly specific and sensitive antigen for SARS-CoV antibody detection.
- To develop a robust serological assay for SARS diagnosis.
Main Methods:
- Expression of SARS-CoV structural proteins as fusion proteins in E. coli.
- Antigenic screening using a protein microarray with sera from SARS patients.
- Development and validation of an indirect antibody ELISA using the identified antigen.
Main Results:
- Recombinant GST-N2 fusion protein (carboxy-terminus of N protein) showed the strongest reactivity and lowest non-specific binding.
- ELISA using GST-N2 detected SARS antibodies in 70.4% of inpatient and 75.8% of convalescent sera.
- A very low false positive rate of 0.07% was observed in non-SARS individuals.
- Protein microarray demonstrated a higher positive rate (86.1%) and sensitivity (1:200 dilution) compared to ELISA.
Conclusions:
- GST-N2 fusion protein is a promising diagnostic antigen for SARS-CoV serological assays.
- Protein microarrays offer a high-throughput platform for rapid antigen screening and SARS diagnosis.
- The developed assay exhibits high sensitivity and specificity for SARS antibody detection.