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
de Zh
| WPRIM
| ID: wpr-286047
Bibliothèque responsable:
WPRO
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.
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Fragments peptidiques
/
Sang
/
Protéines recombinantes
/
Tests sérologiques
/
Chimie
/
Technique de Western
/
Coronavirus
/
Protéines nucléocapside
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Allergie et immunologie
/
Génétique
Type d'étude:
Diagnostic_studies
Limites du sujet:
Adult
/
Humans
langue:
Zh
Texte intégral:
Chinese Journal of Virology
Année:
2011
Type:
Article