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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 Dans Chinois | WPRIM | ID: wpr-305278
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.
Sujets)
Texte intégral: Disponible Indice: WPRIM (Pacifique occidental) Sujet Principal: Virologie / Protéines recombinantes / Test ELISA / Clonage moléculaire / Glycoprotéine hémagglutinine du virus influenza / Diagnostic / Escherichia coli / Grippe humaine / Sous-type H9N2 du virus de la grippe A / Génétique Type d'étude: Etude diagnostique Limites du sujet: Humains langue: Chinois Texte intégral: Chinese Journal of Biotechnology Année: 2005 Type: Article

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Texte intégral: Disponible Indice: WPRIM (Pacifique occidental) Sujet Principal: Virologie / Protéines recombinantes / Test ELISA / Clonage moléculaire / Glycoprotéine hémagglutinine du virus influenza / Diagnostic / Escherichia coli / Grippe humaine / Sous-type H9N2 du virus de la grippe A / Génétique Type d'étude: Etude diagnostique Limites du sujet: Humains langue: Chinois Texte intégral: Chinese Journal of Biotechnology Année: 2005 Type: Article