Pereira S R, Travassos C E, Huguenim A, Guimarães A C, Silva A G, Guimarães M A
Departamento de Virologia, Instituto de Microbiologia Prof. Paulo de Góes, Universidade Federal do Rio de Janeiro, Brasil.
Braz J Med Biol Res. 1998 May;31(5):671-4. doi: 10.1590/s0100-879x1998000500011.
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.
为评估蛋白质印迹法用于诊断传染性法氏囊病病毒（IBDV）感染的效果，用IBDV毒株对鸡进行实验性感染，并通过阻断蛋白质印迹试验（bWB）检测病毒抗原和抗体的存在情况。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）后，将从法氏囊（BF）获得的病毒蛋白转移至硝酸纤维素膜上，并用心脏穿刺采集的鸡血清检测这些蛋白。为消除非特异性反应，我们使用了兔抗鸡血清（阻断剂）。通过bWB试验，检测到了43 kDa（VP2）和32 kDa（VP3）两种不同的病毒蛋白。我们建议将该方法用于检测疑似IBDV再感染及慢性亚临床型疾病的动物的IBDV感染情况。使用兔抗鸡血清进行阻断，该方法实用、灵敏且耗时较少。
