Expression of hemagglutinin of avian influenza virus (AIV) and its application in diagnosis of AIV H9 subtype / 生物工程学报
Chinese Journal of Biotechnology
; (12): 315-319, 2005.
Article
en Zh
| WPRIM
| ID: wpr-305278
Biblioteca responsable:
WPRO
ABSTRACT
In order to differently diagnose avian influenza virus (AIV) subtypes, the HA gene of AIV H9 subtype was cloned, expressed and utilized in an enzyme-linked immunoad sorbent assay (ELISA). HA gene (1683bp) of H9N2 AIV was amplified by RT-PCR from a strain of field isolated H9N2 AIV, and its identity was confirmed by sequencing. The HA gene was subcloned into prokaryotic expression vector pGEX-KG with its secretion signal sequence removed. The expressed HA-GST fusion protein in E. coli BL21 was characterized by SDS-PAGE and western blotting analysis as a 90kD protein with immunogenicity. The fusion protein was present primarily in inclusion bodies and was purified via denaturation and renenaturation. The HA-GST fusion protein was used to establish an indirect ELISA for the detection of antibodies to H9 subtypes of AIV. The assay has 91.57% specificity to H9 AIV, 92.31% sensitivity and excellent reduplication. It could be used to differently detect antibodies to H9 AIV.
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Virología
/
Proteínas Recombinantes
/
Ensayo de Inmunoadsorción Enzimática
/
Clonación Molecular
/
Glicoproteínas Hemaglutininas del Virus de la Influenza
/
Diagnóstico
/
Escherichia coli
/
Gripe Humana
/
Subtipo H9N2 del Virus de la Influenza A
/
Genética
Tipo de estudio:
Diagnostic_studies
Límite:
Humans
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Año:
2005
Tipo del documento:
Article