Western blot detection of infectious bursal disease virus infection.
In order to evaluate the use of a Western blot methodology for the diagnosis of infectious bursal disease virus (IBDV) infection, chickens were experimentally infected with IBDV strains and tested for the presence of viral antigens and antibodies by a blocking Western blot test (bWB). The viral proteins obtained from the bursa of Fabricius (BF) were transferred to a nitrocellulose membrane after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the chicken sera obtained by heart puncture were used for the detection of these proteins. In order to eliminate nonspecific reactions, we used a rabbit anti-chicken serum (blocking tool). By the use of the bWB test, two distinct viral proteins of 43-kDa (VP2) and 32-kDa (VP3) were detected. We suggest the use of this methodology for the detection of IBDV infection in animals suspected of having IBDV reinfection and a chronic subclinical form of the disease. With the use of the rabbit anti-chicken sera for blocking, this method is practical, sensitive and less time consuming.
Departamento de Virologia, Instituto de Microbiologia Prof. Paulo de Góes, Universidade Federal do Rio de Janeiro, Brasil.
S R
SR
Pereira
C E
CE
Travassos
A
A
Huguenim
A C
AC
Guimarães
A G
AG
Silva
M A
MA
Guimarães
D000818:Animals; D000914:Antibodies, Viral / Q000302:isolation & purification; D000956:Antigens, Viral / Q000302:isolation & purification; D018175:Birnaviridae Infections / Q000175:diagnosis*; D015153:Blotting, Western; D002645:Chickens / Q000821:virology*; D007243:Infectious bursal disease virus / Q000276:immunology / Q000302:isolation & purification*; D011201:Poultry Diseases / Q000175:diagnosis*; D014763:Viral Matrix Proteins / Q000302:isolation & purification
D000914:Antibodies, Viral; D000956:Antigens, Viral; D014763:Viral Matrix Proteins