Division of Avian Infectious Diseases, States Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, 427 Maduan Street, Harbin 150001, People's Republic of China.
Arch Virol. 2012 May;157(5):969-73. doi: 10.1007/s00705-012-1256-4. Epub 2012 Feb 25.
Reverse genetic systems for efficient generation of very virulent infectious bursal disease virus (vvIBDV) are currently limited. In this study, we have developed a simple and efficient way to rescue vvIBDV using SPF chickens. The genome of a vvIBDV strain, HLJ0504, flanked by hammerhead and hepatitis delta ribozyme sequences, was cloned downstream of the cytomegalovirus enhancer and the chicken beta-actin promoter of the vector pCAGGS. After transfection of DF-1 cells, cell suspensions were injected into the bursa organ of three-week-old SPF chickens. Using this system, vvIBDV was recovered at high titers after one passage, and the rescued vvIBDV remained highly lethal to SPF chickens. This simple and efficient method to rescue vvIBDV will be a valuable tool for better understanding the molecular virulence determinants of vvIBDV.
目前，用于高效产生非常强毒传染性法氏囊病病毒（vvIBDV）的反向遗传系统受到限制。在本研究中，我们开发了一种使用 SPF 鸡来拯救 vvIBDV 的简单有效的方法。vvIBDV 株 HLJ0504 的基因组，侧翼为锤头和肝炎 delta 核酶序列，被克隆到载体 pCAGGS 的巨细胞病毒增强子和鸡β-肌动蛋白启动子的下游。在 DF-1 细胞中转染后，细胞悬浮液被注入到三周龄 SPF 鸡的法氏囊中。使用该系统，vvIBDV 在一次传代后以高滴度回收，并且拯救的 vvIBDV 仍然对 SPF 鸡具有高度致死性。这种简单有效的拯救 vvIBDV 的方法将成为更好地了解 vvIBDV 分子毒力决定因素的有价值的工具。
