Western blot detection of infectious bursal disease virus infection

Abstract

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. Correspondence

Vaccine development through terminal deletions of an infectious bursal disease virus protein 2 precursor variant

“…Bursas were removed and the chickens' bursas to body weight ratios (BBWR) were calculated. A 10% bursal homogenate in PBS was prepared from the bursal tissue and the homogenates were analyzed using Western blotting analysis to determine whether the challenged chickens were infected by vvIBDV [33].…”

Concentration of Sindbis virus with optimized gradient insulator-based dielectrophoresis

“…Serological methods based on antibody-antigen recognition are a mainstay of viral diagnosis today and include immunofluorescence techniques, [11][12][13] enzyme linked immunosorbent assay, [14][15][16] western blot, [17][18][19] green fluorescent protein, 20 and a host of other less common approaches. Nucleic acid-based assays that rely on polymerase chain reaction (PCR) methods are commonly used both for identification and quantification of viruses.…”