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OJVRTM
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
Microbiology,
ABSTRACT
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