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.
Highlights
A total of 28 chickens were inoculated with infectious bursal disease virus (IBDV)
Chickens inoculated with the Lukert strain (LUK) strain did not show any clinical symptons, animals inoculated with the virulent standard-challenge strain (ST-C) developed signs of illness from the 3rd to the 7th day after inoculation (a)
The use of a rabbit anti-chicken bursa of Fabricius (BF) serum eliminated nonspecific reactions (Figure 1, lanes 1, 2 and 3) due to autoantibodies that are commonly detected in chicken serum infected by IBDV and Marek’s disease virus [14,15,16,17,18,19], which impair the interpretation of the test
Summary
A total of 28 chickens were inoculated with IBDV. Half of them were inoculated with the ST-C strain (virulent) and the other half with the LUK strain (live attenuated). Analysis of IBDV infection by the blocking Western blot (bWB) showed the presence of viral proteins (VP2 and VP4) from the 6th to the 30th day after inoculation.
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Schedule a callMore From: Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas
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