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
in Zh
| WPRIM
| ID: wpr-305278
Responsible library:
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.
Full text:
1
Database:
WPRIM
Main subject:
Virology
/
Recombinant Proteins
/
Enzyme-Linked Immunosorbent Assay
/
Cloning, Molecular
/
Hemagglutinin Glycoproteins, Influenza Virus
/
Diagnosis
/
Escherichia coli
/
Influenza, Human
/
Influenza A Virus, H9N2 Subtype
/
Genetics
Type of study:
Diagnostic_studies
Limits:
Humans
Language:
Zh
Journal:
Chinese Journal of Biotechnology
Year:
2005
Document type:
Article