Expression and purification of different segments from HCoV-NL63 nucleocapsid protein and their application in detection of antibodies / 病毒学报
Chinese Journal of Virology
;
(6): 244-249, 2011.
Article
in Chinese
| WPRIM
| ID: wpr-286047
ABSTRACT
Prokaryotic expression plasmids carrying N-terminal(1-163aa) and C-terminal(141-306aa) gene of HCoV-NL63 nucleocapsid protein were constructed with pET-30a(+) vector. Consequently, we prepared two purified proteins, Np and Cp, respectively, and established a Western blotting-based line assay (WBLA) for detection of antibodies against HCoV-NL63 using three purified proteins Np , Cp and Nf, a full-length HCoV-NL63 nucleocapsid protein as previously reported. We detected anti-HCoV-NL63 antibodies among 50 sera samples collected from adult for health-examination by WBLA. The results showed that 25 (50%), 27 (54%), 36 (72%) of 50 sera were indentified as anti-HCoV-NL63 antibody positive when the antigen was from Nf, Np and Cp, respectively. Among these sera with positive anti-HCoV-NL63 antibody,Cp showed highest antibody positive rate in WBLA,and consistent rates of detection were 64% between Nf and Np, 54% between Nf and Cp, 54% between Np and Cp. Our study provides the foundation for development of HCoV-NL63 serological detection reagents and an experimental tool for immunological research of HCoV-NL63 infection.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Peptide Fragments
/
Blood
/
Recombinant Proteins
/
Serologic Tests
/
Chemistry
/
Blotting, Western
/
Coronavirus
/
Nucleocapsid Proteins
/
Allergy and Immunology
/
Genetics
Type of study:
Diagnostic study
Limits:
Adult
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Virology
Year:
2011
Type:
Article
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