Original scientific paper

Rapid latex agglutination test for serodiagnosis of fowl adenovirus serotype 4 using recombinant antigen

Govindan Kalaiselvi ; Department of Animal Biotechnology, Madras Veterinary College, India
Manoharan Parthiban ; Department of Animal Biotechnology, Madras Veterinary College, India
Madaboosi S. Narayanan ; Department of Animal Biotechnology, Madras Veterinary College, India
Srinivasan S. Kumar ; Department of Animal Biotechnology, Madras Veterinary College, India
Kathaperumal Kumanan ; Department of Animal Biotechnology, Madras Veterinary College, India

Abstract

Hydropericardium hepatitis syndrome (HPS) is a newly emerging disease of poultry, which is caused by fowl adenovirus serotype 4. The virus was propagated in a primary chicken embryo liver culture. The cytopathic effect was observed from the third passage onwards. DNA was isolated from the infected culture and used as a template for amplification of a partial hexon gene (700 bp) using hexon gene specific primers. The amplified product was cloned into pProEX HT b vector and the ligated product was transformed into DH5α cells. The
recombinant clones were analyzed by colony PCR and plasmid isolation, followed by restriction digestion to check the insert release. The positive clones were induced by IPTG. The induced culture fractions were checked at different hours and the induction was high at the 4th hour onwards. The expressed proteins were purified and confirmed by using hyperimmune serum against FAV4 by western blot analysis and the protein size of 50kda was obtained. The purified recombinant FAV4 protein was used as a serodiagnostic agent using enzyme linked immunosorbent assay and latex agglutination test.

Keywords

fowl adenovirus; hexon gene; cloning; expression in E. coli; ELISA; latex agglutination test

Hrčak ID:

62390

URI

https://hrcak.srce.hr/62390

Publication date:

16.12.2010.

Article data in other languages: croatian

Visits: 1.662 *