Production of cell culture based vaccine and DNA Recombinant vaccine against Hydro Pericardium Syndrome (HPS) virus of poultry and their comparison through ELISA.

Production of cell culture based vaccine and DNA Recombinant vaccine against Hydro Pericardium Syndrome (HPS) virus of poultry and their comparison through ELISA Research and Research and Development[.]

Research and Development on Hydro Pericardium Syndrome virus vaccine M Salah-ud-Din Shah, DVM, M.Sc (Hons) Biological Chemistry Division, NIAB INTRODUCTION • Hydropericardium Syndrome is a viral disease of poultry caused by Avi Adeno virus- (Rabbani et al., 1998) • It was first observed in 1987 at Angara goth, a broiler growing area near Karachi, Pakistan So the syndrome was named as Angara disease (Jaffery, 1988) • This disease causes heavy losses every year, mortality rate is 50-70% • Vaccine available in market is 25% of total population (170 M) STATUS OF AVAILABLE VACCINES • At present vaccines available in market are produced from crude methods e.g formalized liver homogenate, these vaccines have following short comings • They carry proteins of livers • Pathogens other than adeno virus if birds are infected • These vaccines have very short shelf life (only 2-3 months) • Vaccine failure is very common PREVIOUS WORK • Vaccine is Produced from crude method with slight modifications • Virus is extracted & purified by centrifugation • Contains no liver protein • Monitoring of chicks for any other disease to avoid other pathogens • Addition of antibiotics to increase shelf life for one year • Addition of Vitamin E and Selenium Marker 116 kDa 66.2 kDa 45 kDa 35 kDa 25 kDa 14.4 kDa 18.2 kDa No of Daily Mortalities in infected flock after vaccination Weakly Antibody titer of broilers after vaccination in treated & control group Brochure of HPS Vaccine PLAN OF WORK • Following techniques can be used for production of standard vaccine • Cell culture based vaccine production • DNA recombinant vaccine production • Trials of both vaccines and comparison through ELISA CELL CULTURE BASED VACCINE • This virus can be grown on chicken hepatocytes cultured in vitro by following steps (Hess et al 1998) • Culturing of primary chicken hepatocytes in cell culture media (M - 199) to obtain monolayer in cell culture flasks • Inoculation of adeno virus on monolayer of cells and allow the virus to grow on cells • Extraction of virus from cells after observing cytopathic effects, quantification by ELISA, then use this virus as antigen in vaccine DNA RECOMBINANT VACCINE • Extraction of viral DNA from samples of • • • • infected chicks (Kao et al 2000) Amplification of HPS virus surface antigen gene by PCR (Jiang et al.,1999) Ligate HPS virus surface antigen gene in cloning vector Transform ligated Product in E coli Screen and select the recombined clone • Confirm the clone by running the sample on 1% agarose gel • DNA sequencing • Detect the cloned protein by ELISA • Selected cloned colony will be used for antigen in vaccine production EVALUATION OF VACCINES • Trials of both vaccines will be done by vaccinating the birds first on small scale then on large scale in field conditions • Collection of serum samples from vaccinated birds • ELISA will be performed at end for determination of antibody titers to evaluate both vaccines (Dawson et al., 1998) Thanks