Johne Reimar, Raue Rüdiger, Grund Christian, Kaleta Erhard F, Müller Hermann
Institute for Virology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 29, D-04103 Leipzig, Germany.
Avian Pathol. 2004 Jun;33(3):328-36. doi: 10.1080/0307945042000220589.
Beak and feather disease virus (BFDV) causes severe disease characterized by irreversible feather disorders and severe immunosuppression in many psittacine species. BFDV cannot be propagated in tissue or cell cultures, rendering virus propagation and thus diagnosis rather difficult. To develop reliable diagnostic methods, the region encoding the BFDV capsid protein C1 was cloned from an infected sulphur-crested cockatoo (Cacatua galerita). Phylogenetic analysis showed this gene had 76.3 to 83.2% amino acid identity to published sequences. No protein was detected after induction of full-length C1 expression in Escherichia coli. However, deletion of an amino-terminal arginine-rich sequence facilitated expression. C1(39-244)-His, a polyhistidine-tailed variant of this protein, was purified and used for immunization of chickens. The immune sera detected C1 with an apparent molecular weight of 27 kDa in western blots of organ homogenates of BFDV-infected birds. Using C1(39-244)-His as antigen, 11 psittacine sera were tested for the presence of BFDV-specific antibodies by enzyme-linked immunosorbent assay and immunoblotting. The results obtained correlated well with the BFDV-specific haemagglutination inhibition activity of the sera, suggesting C1(39-244)-His has value as a recombinant antigen for BFDV-specific serological tests.
喙羽病病毒(BFDV)可引发严重疾病,其特征为许多鹦鹉种类出现不可逆的羽毛紊乱和严重免疫抑制。BFDV无法在组织或细胞培养物中增殖,这使得病毒增殖以及诊断都相当困难。为开发可靠的诊断方法,从一只受感染的硫冠凤头鹦鹉(白凤头鹦鹉)中克隆了编码BFDV衣壳蛋白C1的区域。系统发育分析表明,该基因与已发表序列的氨基酸同一性为76.3%至83.2%。在大肠杆菌中诱导全长C1表达后未检测到蛋白质。然而,删除氨基末端富含精氨酸的序列有助于表达。C1(39 - 244)-His,即该蛋白的多组氨酸尾变体,被纯化并用于免疫鸡。免疫血清在BFDV感染鸟类的器官匀浆的蛋白质印迹中检测到表观分子量为27 kDa的C1。以C1(39 - 244)-His作为抗原,通过酶联免疫吸附测定和免疫印迹检测了11份鹦鹉血清中BFDV特异性抗体的存在情况。所得结果与血清的BFDV特异性血凝抑制活性相关性良好,表明C1(39 -
244)-His作为BFDV特异性血清学检测的重组抗原有一定价值。