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Online Journal of Veterinary Research©
Volume 9 (1):12-18, 2005
Isolation of Egg Drop Syndrome Virus and Protein Fractionation Analysis using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis
Rasool MH, Rahman SU, Mansoor MK, Siddique M
Molecular Virology Laboratory Department of Veterinary
Rasool MH, Rahman SU, Mansoor MK, Siddique M., Isolation of Egg Drop Syndrome Virus and Protein Fractionation Analysis using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis, Online J Vet Res 9 (1):12-18, 2005. Six isolates of egg drop syndrome (EDS) virus were recovered from 5 outbreaks in commercial laying hens. Aberrant eggs were fed to susceptible laying hens to induce infection, and tissue samples were inoculated in 11 days old duck embryos. Virus in harvested allanto-amniotic fluid was verified by spot and microhaemagglutination inhibition and agar gel precipitation tests. Virus grew in duck embryo and agglutinated avian but not mammalian red blood cells. Isolates were purified by velocity density gradient centrifugation and protein concentration was determined by the Lowry method. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was conducted by loading 300µg protein concentration on 12.5% gel using discontinuous buffer system. The six isolates showed 13 polypeptides, which were identical to those described in the referral EDS-76 virus (strain-127). The molecular weights of the polypeptides ranged from 6.5 KDa to 126 KDa.
KEY WORDS: Egg Drop Syndrome, Haemagglutination, Oviduct, Duck embryos, Sodium dodecyl sulphate polyacrylamide gel electrophoresis, polypeptides